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  1. Actinobacillus pleuropneumoniae Possesses an Antiviral Activity against Porcine Reproductive and Respiratory Syndrome Virus

    PubMed Central

    Labrie, Josée; Hernandez Reyes, Yenney; Burciaga Nava, Jorge A.; Gagnon, Carl A.; Jacques, Mario

    2014-01-01

    Pigs are often colonized by more than one bacterial and/or viral species during respiratory tract infections. This phenomenon is known as the porcine respiratory disease complex (PRDC). Actinobacillus pleuropneumoniae (App) and porcine reproductive and respiratory syndrome virus (PRRSV) are pathogens that are frequently involved in PRDC. The main objective of this project was to study the in vitro interactions between these two pathogens and the host cells in the context of mixed infections. To fulfill this objective, PRRSV permissive cell lines such as MARC-145, SJPL, and porcine alveolar macrophages (PAM) were used. A pre-infection with PRRSV was performed at 0.5 multiplicity of infection (MOI) followed by an infection with App at 10 MOI. Bacterial adherence and cell death were compared. Results showed that PRRSV pre-infection did not affect bacterial adherence to the cells. PRRSV and App co-infection produced an additive cytotoxicity effect. Interestingly, a pre-infection of SJPL and PAM cells with App blocked completely PRRSV infection. Incubation of SJPL and PAM cells with an App cell-free culture supernatant is also sufficient to significantly block PRRSV infection. This antiviral activity is not due to LPS but rather by small molecular weight, heat-resistant App metabolites (<1 kDa). The antiviral activity was also observed in SJPL cells infected with swine influenza virus but to a much lower extent compared to PRRSV. More importantly, the PRRSV antiviral activity of App was also seen with PAM, the cells targeted by the virus in vivo during infection in pigs. The antiviral activity might be due, at least in part, to the production of interferon ?. The use of in vitro experimental models to study viral and bacterial co-infections will lead to a better understanding of the interactions between pathogens and their host cells, and could allow the development of novel prophylactic and therapeutic tools. PMID:24878741

  2. Actinobacillus pleuropneumoniae possesses an antiviral activity against porcine reproductive and respiratory syndrome virus.

    PubMed

    Lévesque, Cynthia; Provost, Chantale; Labrie, Josée; Hernandez Reyes, Yenney; Burciaga Nava, Jorge A; Gagnon, Carl A; Jacques, Mario

    2014-01-01

    Pigs are often colonized by more than one bacterial and/or viral species during respiratory tract infections. This phenomenon is known as the porcine respiratory disease complex (PRDC). Actinobacillus pleuropneumoniae (App) and porcine reproductive and respiratory syndrome virus (PRRSV) are pathogens that are frequently involved in PRDC. The main objective of this project was to study the in vitro interactions between these two pathogens and the host cells in the context of mixed infections. To fulfill this objective, PRRSV permissive cell lines such as MARC-145, SJPL, and porcine alveolar macrophages (PAM) were used. A pre-infection with PRRSV was performed at 0.5 multiplicity of infection (MOI) followed by an infection with App at 10 MOI. Bacterial adherence and cell death were compared. Results showed that PRRSV pre-infection did not affect bacterial adherence to the cells. PRRSV and App co-infection produced an additive cytotoxicity effect. Interestingly, a pre-infection of SJPL and PAM cells with App blocked completely PRRSV infection. Incubation of SJPL and PAM cells with an App cell-free culture supernatant is also sufficient to significantly block PRRSV infection. This antiviral activity is not due to LPS but rather by small molecular weight, heat-resistant App metabolites (<1 kDa). The antiviral activity was also observed in SJPL cells infected with swine influenza virus but to a much lower extent compared to PRRSV. More importantly, the PRRSV antiviral activity of App was also seen with PAM, the cells targeted by the virus in vivo during infection in pigs. The antiviral activity might be due, at least in part, to the production of interferon ?. The use of in vitro experimental models to study viral and bacterial co-infections will lead to a better understanding of the interactions between pathogens and their host cells, and could allow the development of novel prophylactic and therapeutic tools. PMID:24878741

  3. The small envelope protein of porcine reproductive and respiratory syndrome virus possesses ion channel protein-like properties

    SciTech Connect

    Lee, Changhee; Yoo, Dongwan . E-mail: dyoo@uoguelph.ca

    2006-11-10

    The small envelope (E) protein of porcine reproductive and respiratory syndrome virus (PRRSV) is a hydrophobic 73 amino acid protein encoded in the internal open reading frame (ORF) of the bicistronic mRNA2. As a first step towards understanding the biological role of E protein during PRRSV replication, E gene expression was blocked in a full-length infectious clone by mutating the ATG translational initiation to GTG, such that the full-length mutant genomic clone was unable to synthesize the E protein. DNA transfection of PRRSV-susceptible cells with the E gene knocked-out genomic clone showed the absence of virus infectivity. P129-{delta}E-transfected cells however produced virion particles in the culture supernatant, and these particles contained viral genomic RNA, demonstrating that the E protein is essential for PRRSV infection but dispensable for virion assembly. Electron microscopy suggests that the P129-{delta}E virions assembled in the absence of E had a similar appearance to the wild-type particles. Strand-specific RT-PCR demonstrated that the E protein-negative, non-infectious P129-{delta}E virus particles were able to enter cells but further steps of replication were interrupted. The entry of PRRSV has been suggested to be via receptor-mediated endocytosis, and lysomotropic basic compounds and known ion-channel blocking agents both inhibited PRRSV replication effectively during the uncoating process. The expression of E protein in Escherichia coli-mediated cell growth arrests and increased the membrane permeability. Cross-linking experiments in cells infected with PRRSV or transfected with E gene showed that the E protein was able to form homo-oligomers. Taken together, our data suggest that the PRRSV E protein is likely an ion-channel protein embedded in the viral envelope and facilitates uncoating of virus and release of the genome in the cytoplasm.

  4. Tourette's syndrome: from demonic possession and psychoanalysis to the discovery of gene.

    PubMed

    Germiniani, Francisco M B; Miranda, Anna Paula P; Ferenczy, Peter; Munhoz, Renato P; Teive, Hélio A G

    2012-07-01

    In this paper we make a brief historical review of the hypothesis concerning the etiology of Tourette's syndrome (TS), focusing on varying trends over time: at first, its presumed relation to witchcraft and demonic possessions, followed by the psychoanalytical theory, which attributed TS to a masturbatory equivalent. Then, progressing to modern time, to the immunological theory and finally the advent of genetics and their role in the etiology of TS. PMID:22836463

  5. [The antigenic properties of hepatitis A virus particles possessing different physicochemical characteristics].

    PubMed

    Kusov, Iu Iu; Nastashenko, T A; Kazachkov, Iu A; Donets, M A; Grishina, G K; Berkova, N P; Balaian, M S

    1991-01-01

    A modified enzyme immunoassay based on adsorption of antihepatitis A virus (HAV) IgG-HRPO conjugate and monoclonal antibodies to HAV were used to investigate antigenic differences between mature HAV virions and subviral particles with different buoyant densities in CsCl produced in HAV-infected cells. The mature virions (1.34 g/cm3) appeared to have common antigenic determinants with subviral particles (1.20, 1.27, and 1.30 g/cm3) and possess some additional determinants. Nevertheless, both subviral particles and mature virions induced antibodies capable of neutralizing HAV infectivity in tissue culture. PMID:1654006

  6. Molecular evolution and antigenic variation of European brown hare syndrome virus (EBHSV).

    PubMed

    Lopes, Ana M; Capucci, Lorenzo; Gavier-Widén, Dolores; Le Gall-Reculé, Ghislaine; Brocchi, Emiliana; Barbieri, Ilaria; Quéméner, Agnès; Le Pendu, Jacques; Geoghegan, Jemma L; Holmes, Edward C; Esteves, Pedro J; Abrantes, Joana

    2014-11-01

    European brown hare syndrome virus (EBHSV) is the aetiological agent of European brown hare syndrome (EBHS), a disease affecting Lepus europaeus and Lepus timidus first diagnosed in Sweden in 1980. To characterize EBHSV evolution we studied hare samples collected in Sweden between 1982 and 2008. Our molecular clock dating is compatible with EBHSV emergence in the 1970s. Phylogenetic analysis revealed two lineages: Group A persisted until 1989 when it apparently suffered extinction; Group B emerged in the mid-1980s and contains the most recent strains. Antigenic differences exist between groups, with loss of reactivity of some MAbs over time, which are associated with amino acid substitutions in recognized epitopes. A role for immune selection is also supported by the presence of positively selected codons in exposed regions of the capsid. Hence, EBHSV evolution is characterized by replacement of Group A by Group B viruses, suggesting that the latter possess a selective advantage. PMID:25155199

  7. Malsoor virus, a novel bat phlebovirus, is closely related to severe fever with thrombocytopenia syndrome virus and heartland virus.

    PubMed

    Mourya, D T; Yadav, P D; Basu, A; Shete, A; Patil, D Y; Zawar, D; Majumdar, T D; Kokate, P; Sarkale, P; Raut, C G; Jadhav, S M

    2014-03-01

    During a survey in the year 2010, a novel phlebovirus was isolated from the Rousettus leschenaultii species of bats in western India. The virus was identified by electron microscopy from infected Vero E6 cells. Phylogenic analysis of the complete genome showed its close relation to severe fever with thrombocytopenia syndrome (SFTS) and Heartland viruses, which makes it imperative to further study its natural ecology and potential as a novel emerging zoonotic virus. PMID:24390329

  8. Porcine reproductive and respiratory syndrome virus replication and quasispecies evolution in pigs that lack adaptive immunity.

    PubMed

    Chen, Nanhua; Dekkers, Jack C M; Ewen, Catherine L; Rowland, Raymond R R

    2015-01-01

    The replication of porcine reproductive and respiratory syndrome virus (PRRSV) was studied in a line of pigs possessing a severe combined immunodeficiency (SCID). Real-time RT-PCR revealed a unique course of infection for the SCID group. During the course of infection, viremia was initially significantly lower than normal littermates, but by 21 days was significantly elevated. Deep sequencing of the viral structural genes at days 11 and 21 identified seven amino acid substitutions in both normal and SCID pigs. The most significant change was a W99R substitution in GP2, which was present in the inoculum at a frequency of 35%, but eventually disappeared from all pigs regardless of immune status. Therefore, amino acid substitutions that appear during acute infection are likely the result of the adaptation of the virus to replication in pigs and not immune selection. PMID:25451069

  9. Borna disease virus possesses an NF-?B inhibitory sequence in the nucleoprotein gene

    PubMed Central

    Makino, Akiko; Fujino, Kan; Parrish, Nicholas F.; Honda, Tomoyuki; Tomonaga, Keizo

    2015-01-01

    Borna disease virus (BDV) has a non-segmented, negative-stranded RNA genome and causes persistent infection in many animal species. Previous study has shown that the activation of the I?B kinase (IKK)/NF-?B pathway is reduced by BDV infection even in cells expressing constitutively active mutant IKK. This result suggests that BDV directly interferes with the IKK/NF-?B pathway. To elucidate the mechanism for the inhibition of NF-?B activation by BDV infection, we evaluated the cross-talk between BDV infection and the NF-?B pathway. Using Multiple EM for Motif Elicitation analysis, we found that the nucleoproteins of BDV (BDV-N) and NF-?B1 share a common ankyrin-like motif. When THP1-CD14 cells were pre-treated with the identified peptide, NF-?B activation by Toll-like receptor ligands was suppressed. The 20S proteasome assay showed that BDV-N and BDV-N-derived peptide inhibited the processing of NF-?B1 p105 into p50. Furthermore, immunoprecipitation assays showed that BDV-N interacted with NF-?B1 but not with NF-?B2, which shares no common motif with BDV-N. These results suggest BDV-N inhibits NF-?B1 processing by the 20S proteasome through its ankyrin-like peptide sequence, resulting in the suppression of IKK/NF-?B pathway activation. This inhibitory effect of BDV on the induction of the host innate immunity might provide benefits against persistent BDV infection. PMID:25733193

  10. White spot syndrome virus inactivation study by using gamma irradiation

    NASA Astrophysics Data System (ADS)

    Heidareh, Marzieh; Sedeh, Farahnaz Motamedi; Soltani, Mehdi; Rajabifar, Saeed; Afsharnasab, Mohammad; Dashtiannasab, Aghil

    2014-09-01

    The present study was conducted to investigate the effect of gamma irradiation on white spot syndrome virus (WSSV). White spot syndrome virus is a pathogen of major economic importance in cultured penaeid shrimp industries. White spot disease can cause mortalities reaching 100% within 3-10 days of gross signs appearing. During the period of culture, immunostimulant agents and vaccines may provide potential methods to protect shrimps from opportunistic and pathogenic microrganisms. In this study, firstly, WSSV was isolated from infected shrimp and then multiplied in crayfish. WSSV was purified from the infected crayfish haemolymph by sucrose gradient and confirmed by transmission electron microscopy. In vivo virus titration was performed in shrimp, Penaeus semisulcatus. The LD50 of live virus stock was calculated 10 5.4/mL. Shrimp post-larvae (1-2 g) were treated with gamma-irradiated (different doses) WSSV (100 to 10-4 dilutions) for a period of 10 days. The dose/survival curve for irradiated and un-irradiated WSSV was drawn; the optimum dose range for inactivation of WSSV and unaltered antigenicity was obtained 14-15 kGy. This preliminary information suggests that shrimp appear to benefit from treatment with gammairradiated WSSV especially at 14-15 KGy.

  11. Aminoterminal Amphipathic ?-Helix AH1 of Hepatitis C Virus Nonstructural Protein 4B Possesses a Dual Role in RNA Replication and Virus Production

    PubMed Central

    Gouttenoire, Jérôme; Montserret, Roland; Paul, David; Castillo, Rosa; Meister, Simon; Bartenschlager, Ralf; Penin, François; Moradpour, Darius

    2014-01-01

    Nonstructural protein 4B (NS4B) is a key organizer of hepatitis C virus (HCV) replication complex formation. In concert with other nonstructural proteins, it induces a specific membrane rearrangement, designated as membranous web, which serves as a scaffold for the HCV replicase. The N-terminal part of NS4B comprises a predicted and a structurally resolved amphipathic ?-helix, designated as AH1 and AH2, respectively. Here, we report a detailed structure-function analysis of NS4B AH1. Circular dichroism and nuclear magnetic resonance structural analyses revealed that AH1 folds into an amphipathic ?-helix extending from NS4B amino acid 4 to 32, with positively charged residues flanking the helix. These residues are conserved among hepaciviruses. Mutagenesis and selection of pseudorevertants revealed an important role of these residues in RNA replication by affecting the biogenesis of double-membrane vesicles making up the membranous web. Moreover, alanine substitution of conserved acidic residues on the hydrophilic side of the helix reduced infectivity without significantly affecting RNA replication, indicating that AH1 is also involved in virus production. Selective membrane permeabilization and immunofluorescence microscopy analyses of a functional replicon harboring an epitope tag between NS4B AH1 and AH2 revealed a dual membrane topology of the N-terminal part of NS4B during HCV RNA replication. Luminal translocation was unaffected by the mutations introduced into AH1, but was abrogated by mutations introduced into AH2. In conclusion, our study reports the three-dimensional structure of AH1 from HCV NS4B, and highlights the importance of positively charged amino acid residues flanking this amphipathic ?-helix in membranous web formation and RNA replication. In addition, we demonstrate that AH1 possesses a dual role in RNA replication and virus production, potentially governed by different topologies of the N-terminal part of NS4B. PMID:25392992

  12. Anterior opercular syndrome induced by Epstein-Barr virus encephalitis.

    PubMed

    Matsushima, Takashi; Nishioka, Kenya; Tanaka, Ryota; Yokoyama, Kazumasa; Hattori, Nobutaka

    2016-02-01

    We report a 19-year-old female presenting with fever, drooling, anarthria, and voluntary facial movement disruption, characteristic of anterior opercular syndrome (AOS). Serological examination revealed Epstein-Barr virus (EBV) infection following acute encephalitis with severe ataxia. A single-photon emission computerized tomography (SPECT) examination indicated hypoperfusion in the left perisylvian region, bilateral thalamus, occipital lobe, and cerebellum. This is the first report of AOS related to EBV encephalitis. SPECT was a useful method for detecting the damaged region of the operculum. In addition, AOS is a clinically distinct entity that may help us understand the mechanisms of language circuits within the operculum. PMID:26027635

  13. West Nile Virus Encephalitis Induced Opsoclonus-Myoclonus Syndrome

    PubMed Central

    Cooper, Chad J.; Said, Sarmad

    2014-01-01

    West Nile virus (WNV) is an arthropod borne neurotropic single stranded RNA flavivirus with <1% developing presenting with neurological disease. Immunocompromised and elderly patients are more prone to developing WNV meningitis or encephalitis. Definitive diagnosis of WNV meningoencephalitis is a combination of clinical suspicion and cerebrospinal fluid (CSF) serology. Forty-eight year old Caucasian female presented with a sudden onset of altered mental status after being found unresponsive. She was confused with intermittent bouts of alertness/lethargy and unintelligible responses to questioning. Her medical problems included endometrial cancer that was in remission after undergoing a total abdominal hysterectomy with bilateral salpingectomy and postoperative chemotherapy with paclitaxel and carboplatin. Pertinent physical examination revealed muscle strength that was significantly decreased, nuchal rigidity and +2 pitting edema of both lower extremities. Computed tomography and magnetic resonance imaging of the brain were negative for any intracranial pathology. CSF analysis was consistent with aseptic meningitis with all CSF serology being negative except for positive WNV antibody. A few days after being admitted she developed involuntary random movements of her eyes and generalized jerking movements (myoclonus). This was determined to be opsoclonus myoclonus syndrome (OMS) induced by the WNV meningoencephalitis. She then received five consecutive days of plasmapheresis with a significant improvement in her neurological status. Opsoclonus-myoclonus syndrome (OMS) is a rare neurological disorder associated with chaotic multidirectional eye movements, myoclonus and less frequently cerebellar ataxia. OMS affects as few as 1 in 10,000,000 people per year. The pathogenesis is not fully understood with the majority of cases of opsoclonus-myoclonus syndrome being idiopathic. According to current medical literature there have only been two previous case reports of opsoclonus myoclonus syndrome associated with WNV encephalitis. PMID:24987503

  14. Porites white patch syndrome: associated viruses and disease physiology

    NASA Astrophysics Data System (ADS)

    Lawrence, S. A.; Davy, J. E.; Wilson, W. H.; Hoegh-Guldberg, O.; Davy, S. K.

    2015-03-01

    In recent decades, coral reefs worldwide have undergone significant changes in response to various environmental and anthropogenic impacts. Among the numerous causes of reef degradation, coral disease is one factor that is to a large extent still poorly understood. Here, we characterize the physiology of white patch syndrome (WPS), a disease affecting poritid corals on the Great Barrier Reef. WPS manifests as small, generally discrete patches of tissue discolouration. Physiological analysis revealed that chlorophyll a content was significantly lower in lesions than in healthy tissues, while host protein content remained constant, suggesting that host tissue is not affected by WPS. This was confirmed by transmission electron microscope (TEM) examination, which showed intact host tissue within lesions. TEM also revealed that Symbiodinium cells are lost from the host gastrodermis with no apparent harm caused to the surrounding host tissue. Also present in the electron micrographs were numerous virus-like particles (VLPs), in both coral and Symbiodinium cells. Small (<50 nm diameter) icosahedral VLPs were significantly more abundant in coral tissue taken from diseased colonies, and there was an apparent, but not statistically significant, increase in abundance of filamentous VLPs in Symbiodinium cells from diseased colonies. There was no apparent increase in prokaryotic or eukaryotic microbial abundance in diseased colonies. Taken together, these results suggest that viruses infecting the coral and/or its resident Symbiodinium cells may be the causative agents of WPS.

  15. Severe Fever with Thrombocytopenia Syndrome Virus in Ticks Collected from Humans, South Korea, 2013

    PubMed Central

    Yun, Seok-Min; Lee, Wook-Gyo; Ryou, Jungsang; Yang, Sung-Chan; Park, Sun-Whan; Roh, Jong Yeol; Lee, Ye-Ji; Park, Chan

    2014-01-01

    We investigated the infection rate for severe fever with thrombocytopenia syndrome virus (SFTSV) among ticks collected from humans during May–October 2013 in South Korea. Haemaphysalis longicornis ticks have been considered the SFTSV vector. However, we detected the virus in H. longicornis, Amblyomma testudinarium, and Ixodes nipponensis ticks, indicating additional potential SFTSV vectors. PMID:25061851

  16. Progress in Porcine Respiratory and Reproductive Syndrome Virus Biology and Control

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This special issue of Virus Research is focused on Porcine Reproductive and Respiratory Syndrome (PRRS). It contains 19 reviews invited by CoEditors, Joan K. Lunney and R.R.R. Rowland, on this arterivirus, referred to as PRRS virus (PRRSV), and associated issues. It targets areas such as: phylogenet...

  17. Characterization of the Taura syndrome virus isolate originating from the 2004 Texas Epizootic in cultured shrimp

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Taura syndrome virus (TSV) is a major viral pathogen of penaeid shrimp worldwide. A comprehensive investigation of the Texas isolate of TSV that caused epizootics in shrimp farms in Texas in 2004 (Us04Pv1) revealed that the virus was highly virulent in laboratory bioassays causing severe symptom dev...

  18. Persistence and Retention of Porcine Reproductive and Respiratory Syndrome Virus in Stable Flies (Diptera: Muscidae).

    PubMed

    Rochon, K; Baker, R B; Almond, G W; Gimeno, I M; Pérez de León, A A; Watson, D W

    2015-09-01

    We investigated the acquisition of porcine reproductive and respiratory syndrome (PRRS) virus by the stable fly (Diptera: Muscidae; Stomoxys calcitrans (L.)) through a bloodmeal, and virus persistence in the digestive organs of the fly using virus isolation and quantitative reverse-transcription PCR (qRT-PCR). Stable flies were fed blood containing live virus, modified live vaccine virus, chemically inactivated virus, or no virus. Stable flies acquired PRRSV from the bloodmeal and the amount of virus in the flies declined with time, indicating virus did not replicate in fly digestive tissues. Virus RNA was recovered from the flies fed live virus up to 24?h postfeeding using virus isolation techniques and 96?h using qRT-PCR. We further examined the fate of PRRSV in the hemolymph of the flies following intrathoracic injection to bypass the midgut barrier. PRRSV was detected in intrathoracically inoculated adult stable flies for 10 d using qRT-PCR. In contrast to what we observed in the digestive tract, detectable virus quantities in the intrathoracically inoculated stable flies followed an exponential decay curve. The amount of virus decreased fourfold in the first 3 d and remained stable thereafter, up to 10 d. PMID:26336236

  19. A Plant Extract of Ribes nigrum folium Possesses Anti-Influenza Virus Activity In Vitro and In Vivo by Preventing Virus Entry to Host Cells

    PubMed Central

    Ehrhardt, Christina; Dudek, Sabine Eva; Holzberg, Magdalena; Urban, Sabine; Hrincius, Eike Roman; Haasbach, Emanuel; Seyer, Roman; Lapuse, Julia; Planz, Oliver; Ludwig, Stephan

    2013-01-01

    Infections with influenza A viruses (IAV) are still amongst the major causes of highly contagious severe respiratory diseases not only bearing a devastating effect to human health, but also significantly impact the economy. Besides vaccination that represents the best option to protect from IAV infections, only two classes of anti-influenza drugs, inhibitors of the M2 ion channel and the neuraminidase, often causing resistant IAV variants have been approved. That is why the need for effective and amply available antivirals against IAV is of high priority. Here we introduce LADANIA067 from the leaves of the wild black currant (Ribes nigrum folium) as a potent compound against IAV infections in vitro and in vivo. LADANIA067 treatment resulted in a reduction of progeny virus titers in cell cultures infected with prototype avian and human influenza virus strains of different subtypes. At the effective dose of 100 µg/ml the extract did not exhibit apparent harming effects on cell viability, metabolism or proliferation. Further, viruses showed no tendency to develop resistance to LADANIA067 when compared to amantadine that resulted in the generation of resistant variants after only a few passages. On a molecular basis the protective effect of LADANIA067 appears to be mainly due to interference with virus internalisation. In the mouse infection model LADANIA067 treatment reduces progeny virus titers in the lung upon intranasal application. In conclusion, an extract from the leaves of the wild black currant might be a promising source for the development of new antiviral compounds to fight IAV infections. PMID:23717460

  20. Suppression of shrimp melanization during white spot syndrome virus infection.

    PubMed

    Sutthangkul, Jantiwan; Amparyup, Piti; Charoensapsri, Walaiporn; Senapin, Saengchan; Phiwsaiya, Kornsunee; Tassanakajon, Anchalee

    2015-03-01

    The melanization cascade, activated by the prophenoloxidase (proPO) system, plays a key role in the production of cytotoxic intermediates, as well as melanin products for microbial sequestration in invertebrates. Here, we show that the proPO system is an important component of the Penaeus monodon shrimp immune defense toward a major viral pathogen, white spot syndrome virus (WSSV). Gene silencing of PmproPO(s) resulted in increased cumulative shrimp mortality after WSSV infection, whereas incubation of WSSV with an in vitro melanization reaction prior to injection into shrimp significantly increased the shrimp survival rate. The hemolymph phenoloxidase (PO) activity of WSSV-infected shrimp was extremely reduced at days 2 and 3 post-injection compared with uninfected shrimp but was fully restored after the addition of exogenous trypsin, suggesting that WSSV probably inhibits the activity of some proteinases in the proPO cascade. Using yeast two-hybrid screening and co-immunoprecipitation assays, the viral protein WSSV453 was found to interact with the proPO-activating enzyme 2 (PmPPAE2) of P. monodon. Gene silencing of WSSV453 showed a significant increase of PO activity in WSSV-infected shrimp, whereas co-silencing of WSSV453 and PmPPAE2 did not, suggesting that silencing of WSSV453 partially restored the PO activity via PmPPAE2 in WSSV-infected shrimp. Moreover, the activation of PO activity in shrimp plasma by PmPPAE2 was significantly decreased by preincubation with recombinant WSSV453. These results suggest that the inhibition of the shrimp proPO system by WSSV partly occurs via the PmPPAE2-inhibiting activity of WSSV453. PMID:25572398

  1. Duck Egg-Drop Syndrome Caused by BYD Virus, a New Tembusu-Related Flavivirus

    PubMed Central

    Yu, Xiuling; Wang, Yongyue; Liu, Peipei; Lu, Xishan; Zhang, Guozhong; Hu, Xueying; Liu, Di; Li, Xiaoxia; Su, Wenliang; Lu, Hao; Mok, Ngai Shing; Wang, Peiyi; Wang, Ming; Tian, Kegong; Gao, George F.

    2011-01-01

    Since April 2010, a severe outbreak of duck viral infection, with egg drop, feed uptake decline and ovary-oviduct disease, has spread around the major duck-producing regions in China. A new virus, named BYD virus, was isolated in different areas, and a similar disease was reproduced in healthy egg-producing ducks, infecting with the isolated virus. The virus was re-isolated from the affected ducks and replicated well in primary duck embryo fibroblasts and Vero cells, causing the cytopathic effect. The virus was identified as an enveloped positive-stranded RNA virus with a size of approximately 55 nm in diameter. Genomic sequencing of the isolated virus revealed that it is closely related to Tembusu virus (a mosquito-borne Ntaya group flavivirus), with 87–91% nucleotide identity of the partial E (envelope) proteins to that of Tembusu virus and 72% of the entire genome coding sequence with Bagaza virus, the most closely related flavivirus with an entirely sequenced genome. Collectively our systematic studies fulfill Koch's postulates, and therefore, the causative agent of the duck egg drop syndrome occurring in China is a new flavivirus. Flavivirus is an emerging and re-emerging zoonotic pathogen and BYD virus that causes severe egg-drop, could be disastrous for the duck industry. More importantly its public health concerns should also be evaluated, and its epidemiology should be closely watched due to the zoonotic nature of flaviviruses. PMID:21455312

  2. Influenza A virus and Reye's syndrome in adults.

    PubMed Central

    Davis, L E; Kornfeld, M

    1980-01-01

    We report fatal Reye's syndrome in two adults following proven influenza A viral infections. Reye's syndrome is, therefore, not confined to children but may also occur in adults. Many reported cases of postinfluenza A encephalopathy have clinical and pathological features of Reye's syndrome suggesting that they are not due to postinfectious perivenous demyelination. Images PMID:7205293

  3. Zika virus infection complicated by Guillain-Barre syndrome--case report, French Polynesia, December 2013.

    PubMed

    Oehler, E; Watrin, L; Larre, P; Leparc-Goffart, I; Lastere, S; Valour, F; Baudouin, L; Mallet, Hp; Musso, D; Ghawche, F

    2014-01-01

    Zika fever, considered as an emerging disease of arboviral origin, because of its expanding geographic area, is known as a benign infection usually presenting as an influenza-like illness with cutaneous rash. So far, Zika virus infection has never led to hospitalisation. We describe the first case of Guillain-Barré syndrome (GBS) occurring immediately after a Zika virus infection, during the current Zika and type 1 and 3 dengue fever co-epidemics in French Polynesia. PMID:24626205

  4. A white spot syndrome virus microRNA promotes the virus infection by targeting the host STAT

    PubMed Central

    Ren, Qian; Huang, Ying; He, Yaodong; Wang, Wen; Zhang, Xiaobo

    2015-01-01

    JAK/STAT pathway plays an important role in invertebrates during virus infection. However the microRNA (miRNA)-mediated regulation of JAK/STAT is not intensively investigated. Viral miRNAs, encoded by virus genome, have emerged as important regulators in the virus-host interactions. In this study, a WSSV (white spot syndrome virus)-encoded miRNA (WSSV-miR-22) was characterized in shrimp during virus infection. The results showed that the viral miRNA could promote WSSV infection in shrimp by targeting the host STAT gene. When the expression of JAK or STAT was knocked down by sequence-specific siRNA, the WSSV copies in shrimp were significantly increased, indicating that the JAK/STAT played positive roles in the antiviral immunity of shrimp. The further findings revealed that TEP1 and TEP2 were the effectors of JAK-STAT signaling pathway. The silencing of TEP1 or TEP2 led to an increase of WSSV copies in shrimp, showing TEP1 and TEP2 were involved in the shrimp immune response against virus infection. Therefore our study presented a novel viral miRNA-mediated JAK/STAT-TEP1/TEP2 signaling pathway in virus infection. PMID:26671453

  5. A white spot syndrome virus microRNA promotes the virus infection by targeting the host STAT.

    PubMed

    Ren, Qian; Huang, Ying; He, Yaodong; Wang, Wen; Zhang, Xiaobo

    2015-01-01

    JAK/STAT pathway plays an important role in invertebrates during virus infection. However the microRNA (miRNA)-mediated regulation of JAK/STAT is not intensively investigated. Viral miRNAs, encoded by virus genome, have emerged as important regulators in the virus-host interactions. In this study, a WSSV (white spot syndrome virus)-encoded miRNA (WSSV-miR-22) was characterized in shrimp during virus infection. The results showed that the viral miRNA could promote WSSV infection in shrimp by targeting the host STAT gene. When the expression of JAK or STAT was knocked down by sequence-specific siRNA, the WSSV copies in shrimp were significantly increased, indicating that the JAK/STAT played positive roles in the antiviral immunity of shrimp. The further findings revealed that TEP1 and TEP2 were the effectors of JAK-STAT signaling pathway. The silencing of TEP1 or TEP2 led to an increase of WSSV copies in shrimp, showing TEP1 and TEP2 were involved in the shrimp immune response against virus infection. Therefore our study presented a novel viral miRNA-mediated JAK/STAT-TEP1/TEP2 signaling pathway in virus infection. PMID:26671453

  6. Highly pathogenic Chinese porcine reproductive and respiratory syndrome virus strain JXwn06 in US swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 2006, a large-scale outbreak of highly pathogenic atypical porcine reproductive and respiratory syndrome virus (PRRSV) spread throughout the swine population in China. Causative PRRSV isolates were characterized genetically by a unique 30aa deletion in PRRSV nonstructural protein 2 and clinically...

  7. Pathogenicity and Molecular Characterization of Emerging Porcine Reproductive and Respiratory Syndrome Virus in Vietnam in 2007

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 2007, Vietnam experienced swine disease outbreaks causing clinical signs similar to the "porcine high fever disease" that occurred in China during 2006. Analysis of diagnostic samples from the disease outbreaks in Vietnam identified porcine reproductive and respiratory syndrome virus (PRRSV) and ...

  8. GENETIC CONTROL OF SWINE RESPONSES TO PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS INFECTION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our goal is use the Swine Protein-Annotated Oligonucleotide Microarray (www.pigoligoarray.org) to identify immune regulatory and protective pathways to uncover genetic components involved in early immune responses during porcine reproductive and respiratory syndrome virus (PRRSV) infection. Animals ...

  9. Induction of type I interferons by a novel porcine reproductive and respiratory syndrome virus isolate

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome virus (PRRSV) inhibits synthesis of type I interferons (IFNs) in infected pigs and in cultured cells. Here we report that one PRRSV mutant A2MC2 induces type I IFNs in cultured cells and has no effect on IFN downstream signaling. The mutant isolate was p...

  10. Inhibition of porcine reproductive and respiratory syndrome virus in vitro by forsythoside A

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome virus (PRRSV) represents a significant challenge to the swine industry worldwide. Current control strategies against PRRSV are still inadequate and there is a need for new antiviral therapy method. Forsythoside is a compound derived from fruit of Forsy...

  11. GENETIC CONTROL OF SWINE RESPONSES TO PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS INFECTION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our goal is to uncover genetic components involved in early immune responses during porcine reproductive and respiratory syndrome virus (PRRSV) infection. PRRS costs U.S. swine producers >$700 million annually. We want to determine what are the most significant pathways and genes involved in early i...

  12. Proteolytic Products of the Porcine Reproductive and Respiratory Syndrome Virus Nsp2 Replicase Protein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The nsp2 replicase protein of porcine reproductive and respiratory syndrome virus (PRRSV) was recently demonstrated to be processed from its precursor by the PL2 protease at or near the G1196|G1197 dipeptide in transfected CHO cells. Here, the proteolytic cleavage of PRRSV nsp2 was further investiga...

  13. Proteolytic processing of Porcine Reproductive and Respiratory Syndrome Virus nsp2 replicase protein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One critical step in porcine reproductive and respiratory syndrome virus (PRRSV) replication is the proteolytic processing of the ORF1 polyprotein (replicase). The replicase polyprotein is generally believed to be processed to generate at least 12 smaller nonstructural proteins (nsps) involved in r...

  14. Vaccine efficacy and immune response to swine influenza virus challenge in pigs infected with porcine reproductive and respiratory syndrome virus at the time of SIV-vaccination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to assess the effect of concurrent infection with porcine reproductive and respiratory syndrome virus (PRRSV) on the efficacy of an inactivated swine influenza virus (SIV) vaccine. Eight groups of pigs were used in the study. One group was infected with a virulent PR...

  15. [Study on using NSP2 protein of porcine reproductive and respiratory syndrome virus (HuN4-F112) to express E2 neutralizing epitope of classical swine fever virus].

    PubMed

    Xu, Yan-Zhao; Zhou, Yan-Jun; Tong, Wu; Li, Ling; Jiang, Yi-Feng; Tong, Guang-Zhi

    2013-01-01

    Establishment of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) with co-expression E2 Epitope of Classical Swine Fever virus (CSFV) is a crucial step to develop a genetic engineered vaccine against PRRSV and CSFV. Reverse genetic manipulation could be adopted as a com monly used technique. In this study, we focus on using nonessential regions of NSP2 (aa480-532 and aa508-532) as viral vector to express E2 Epitope of CSFV. A neutralizing epitope of classical swine fever virus (CSFV) E2 protein was inserted into the two nonessential region of nsp2 by the method of mutant PCR, basing on the infectious clone of HuN4-F112 vaccine strain. The co-expressed full-length cDNA clones (psk-HuN4-F112-delta508-532 + E2 and psk-HuN4-F112-delta480-532 + E2) were assembled by cloning and splice of the gene fragments. The completely assembled full-length cDNA clones were confirmed by sequence and Swa I enzyme digestion. Capped RNAs were transcribed in vitro from a full-length cDNA clone of the viral genome and transfected into BHK-21 cells by liposome to acquire the rescued virus. The rescued recombinant viruses were passaged on MARC-145 cells. The successfully rescued viruses were tested by RT-PCR, digestion, and genome sequence. The results showed that these rescued viruses could be distinguished from the parental virus (HuN4-F112) with the mutant genetic marker (Mlu I enzyme site of virual genome at 14 667nt was created by synonymous mutation) and the inserted nsp2 gene region. The results of IFA showed that the inserted E2 epitope could be expressed by the recombinant viruses and the E2 epitope gene was stable during the viral serial passage. The results of plaque assay and viral growth curve showed that the recovery viruses possessed similar characterses of viral growth to those of the parental virus. In summary, the full-length infectious cDNA clones containing the marker gene were constructed and the marker recombinant viruses were rescued. The results suggested that these stable infectious clones could be used as an important tool for development of novel vaccine against PRRSV. PMID:23547375

  16. Plant-Derived Chimeric Virus Particles for the Diagnosis of Primary Sjögren Syndrome

    PubMed Central

    Tinazzi, Elisa; Merlin, Matilde; Bason, Caterina; Beri, Ruggero; Zampieri, Roberta; Lico, Chiara; Bartoloni, Elena; Puccetti, Antonio; Lunardi, Claudio; Pezzotti, Mario; Avesani, Linda

    2015-01-01

    Plants are ideal for the production of protein-based nanomaterials because they synthesize and assemble complex multimeric proteins that cannot be expressed efficiently using other platforms. Plant viruses can be thought of as self-replicating proteinaceous nanomaterials generally stable and easily produced in high titers. We used Potato virus X (PVX), chimeric virus particles, and Cowpea mosaic virus, empty virus-like particles to display a linear peptide (lipo) derived from human lipocalin, which is immunodominant in Sjögren’s syndrome (SjS) and is thus recognized by autoantibodies in SjS patient serum. These virus-derived nanoparticles were thus used to develop a diagnostic assay for SjS based on a direct enzyme linked immunosorbent assay format. We found that PVX-lipo formulations were more sensitive than the chemically synthesized immunodominant peptide and equally specific when used to distinguish between healthy individuals and SjS patients. Our novel assay therefore allows the diagnosis of SjS using a simple, low-invasive serum test, contrasting with the invasive labial biopsy required for current tests. Our results demonstrate that nanomaterials based on plant viruses can be used as diagnostic reagents for SjS, and could also be developed for the diagnosis of other diseases. PMID:26648961

  17. Plant-Derived Chimeric Virus Particles for the Diagnosis of Primary Sjögren Syndrome.

    PubMed

    Tinazzi, Elisa; Merlin, Matilde; Bason, Caterina; Beri, Ruggero; Zampieri, Roberta; Lico, Chiara; Bartoloni, Elena; Puccetti, Antonio; Lunardi, Claudio; Pezzotti, Mario; Avesani, Linda

    2015-01-01

    Plants are ideal for the production of protein-based nanomaterials because they synthesize and assemble complex multimeric proteins that cannot be expressed efficiently using other platforms. Plant viruses can be thought of as self-replicating proteinaceous nanomaterials generally stable and easily produced in high titers. We used Potato virus X (PVX), chimeric virus particles, and Cowpea mosaic virus, empty virus-like particles to display a linear peptide (lipo) derived from human lipocalin, which is immunodominant in Sjögren's syndrome (SjS) and is thus recognized by autoantibodies in SjS patient serum. These virus-derived nanoparticles were thus used to develop a diagnostic assay for SjS based on a direct enzyme linked immunosorbent assay format. We found that PVX-lipo formulations were more sensitive than the chemically synthesized immunodominant peptide and equally specific when used to distinguish between healthy individuals and SjS patients. Our novel assay therefore allows the diagnosis of SjS using a simple, low-invasive serum test, contrasting with the invasive labial biopsy required for current tests. Our results demonstrate that nanomaterials based on plant viruses can be used as diagnostic reagents for SjS, and could also be developed for the diagnosis of other diseases. PMID:26648961

  18. Challenges and opportunities for the control and elimination of porcine reproductive and respiratory syndrome virus.

    PubMed

    Rowland, R R R; Morrison, R B

    2012-03-01

    The control and elimination of porcine reproductive and respiratory syndrome virus (PRRSV) represent two of the most challenging tasks facing the pig industry worldwide. Several factors related to the biology of the virus make disease detection and elimination difficult. Efforts are further hampered by the lack of vaccines that can protect naïve herds from infection. With this in mind, elimination efforts are being initiated which incorporate existing tools and knowledge. A new approach extends herd control strategies to the level of a region. One example of success in PRRSV regional elimination is the Stevens County project in Minnesota. PMID:25471243

  19. Middle East Respiratory Syndrome Coronavirus Spike Protein Delivered by Modified Vaccinia Virus Ankara Efficiently Induces Virus-Neutralizing Antibodies

    PubMed Central

    Song, Fei; Fux, Robert; Provacia, Lisette B.; Volz, Asisa; Eickmann, Markus; Becker, Stephan; Osterhaus, Albert D. M. E.; Haagmans, Bart L.

    2013-01-01

    Middle East respiratory syndrome coronavirus (MERS-CoV) has recently emerged as a causative agent of severe respiratory disease in humans. Here, we constructed recombinant modified vaccinia virus Ankara (MVA) expressing full-length MERS-CoV spike (S) protein (MVA-MERS-S). The genetic stability and growth characteristics of MVA-MERS-S make it a suitable candidate vaccine for clinical testing. Vaccinated mice produced high levels of serum antibodies neutralizing MERS-CoV. Thus, MVA-MERS-S may serve for further development of an emergency vaccine against MERS-CoV. PMID:23986586

  20. Mutations within the nuclear localization signal of the porcine reproductive and respiratory syndrome virus nucleocapsid protein attenuate virus replication

    SciTech Connect

    Lee, Changhee; Hodgins, Douglas; Calvert, Jay G.; Welch, Siao-Kun W.; Jolie, Rika; Yoo, Dongwan . E-mail: dyoo@uoguelph.ca

    2006-03-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is an RNA virus replicating in the cytoplasm, but the nucleocapsid (N) protein is specifically localized to the nucleus and nucleolus in virus-infected cells. A 'pat7' motif of 41-PGKK(N/S)KK has previously been identified in the N protein as the functional nuclear localization signal (NLS); however, the biological consequences of N protein nuclear localization are unknown. In the present study, the role of N protein nuclear localization during infection was investigated in pigs using an NLS-null mutant virus. When two lysines at 43 and 44 at the NLS locus were substituted to glycines, the modified NLS with 41-PGGGNKK restricted the N protein to the cytoplasm. This NLS-null mutation was introduced into a full-length infectious cDNA clone of PRRSV. Upon transfection of cells, the NLS-null full-length clone induced cytopathic effects and produced infectious progeny. The NLS-null virus grew to a titer 100-fold lower than that of wild-type virus. To examine the response to NLS-null PRRSV in the natural host, three groups of pigs, consisting of seven animals per group, were intranasally inoculated with wild-type, placebo, or NLS-null virus, and the animals were maintained for 4 weeks. The NLS-null-infected pigs had a significantly shorter mean duration of viremia than wild-type-infected pigs but developed significantly higher titers of neutralizing antibodies. Mutations occurred at the NLS locus in one pig during viremia, and four types of mutations were identified: 41-PGRGNKK, 41-PGGRNKK, and 41-PGRRNKK, and 41-PGKKSKK. Both wild-type and NLS-null viruses persisted in the tonsils for at least 4 weeks, and the NLS-null virus persisting in the tonsils was found to be mutated to either 41-PGRGNKK or 41-PGGRNKK in all pigs. No other mutation was found in the N gene. All types of reversions which occurred during viremia and persistence were able to translocate the mutated N proteins to the nucleus, indicating a strong selection pressure for reversion at the NLS locus of the N protein in vivo. Reversions from NLS-null to functional NLS in the tonsils suggest a possible correlation of viral persistence with N protein nuclear localization. These results show that N protein nuclear localization is non-essential for PRRSV multiplication but may play an important role in viral attenuation and in pathogenesis in vivo.

  1. Ramsay Hunt syndrome

    MedlinePLUS

    Hunt syndrome; Herpes oticus ... The varicella zoster virus that causes Ramsay Hunt syndrome is the same virus that causes chickenpox and shingles. In people with Ramsay Hunt syndrome, the virus is believed to infect the ...

  2. Infection of United States swine with a Chinese highly pathogenic strain of porcine reproductive and respiratory syndrome virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To assess the pathogenic effects of Type 2 highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) on healthy 10-week old commercial swine in the United States, viral kinetics and resultant disease caused by intranasal inoculation of such virus rescued from an infectious clo...

  3. Associations among Haemophilus parasuis, Mycoplasma hyorhinis, and porcine reproductive and respiratory syndrome virus infections in pigs with polyserositis

    PubMed Central

    Palzer, Andreas; Haedke, Kristina; Heinritzi, Karl; Zoels, Susanne; Ladinig, Andrea; Ritzmann, Mathias

    2015-01-01

    The objective of this study was to determine the associations among Haemophilus parasuis, Mycoplasma hyorhinis, and porcine reproductive and respiratory syndrome (PRRS) virus (EU-field strain) infections in 95 pigs with polyserositis. A significant association between H. parasuis and M. hyorhinis was identified. H. parasuis and M. hyorhinis were significantly more often detected in PRRS virus positive pigs. PMID:25750450

  4. Bayou virus-associated hantavirus pulmonary syndrome in Eastern Texas: identification of the rice rat, Oryzomys palustris, as reservoir host.

    PubMed Central

    Torrez-Martinez, N.; Bharadwaj, M.; Goade, D.; Delury, J.; Moran, P.; Hicks, B.; Nix, B.; Davis, J. L.; Hjelle, B.

    1998-01-01

    We describe the third known case of hantavirus pulmonary syndrome (HPS) due to Bayou virus, from Jefferson County, Texas. By using molecular epidemiologic methods, we show that rice rats (Oryzomys palustris) are frequently infected with Bayou virus and that viral RNA sequences from HPS patients are similar to those from nearby rice rats. Bayou virus is associated with O. palustris; this rodent appears to be its predominant reservoir host. PMID:9452404

  5. Genetic diversity of porcine reproductive and respiratory syndrome virus in Korea.

    PubMed

    Choi, Eun-Jin; Lee, Chang-Hee; Song, Jae-Young; Song, Hee-Jong; Park, Choi-Kyu; Kim, Byounghan; Shin, Yeun-Kyung

    2013-01-01

    The high genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) has been an obstacle to developing an effective vaccine for porcine reproductive and respiratory syndrome (PRRS). This study was performed to assess the degree of genetic diversity among PRRSVs from Korean pig farms where wasting and respiratory syndrome was observed from 2005 to 2009. Samples from 786 farms were tested for the presence of PRRSV using reverse transcription PCR protocol. A total of 117 farms were positive for type 1 PRRSV while 198 farms were positive for type 2. Nucleotide sequences encoding the open reading frame (ORF) 5 were analyzed and compared to those of various published PRRSV isolates obtained worldwide. Sequence identity of the ORF 5 in the isolates was 81.6˜100% for type 1 viruses and 81.4˜100% for type 2 viruses. Phylogenetic analysis of the ORF 5 sequences showed that types 1 and 2 PRRSVs from Korea were mainly classified into three and four clusters, respectively. The analyzed isolates were distributed throughout the clusters independent of the isolation year or geographical origin. In conclusion, our results indicated that the genetic diversity of PRRSVs from Korean pig farms is high and has been increasing over time. PMID:23628658

  6. Pathogenesis of nonsuppurative encephalitis caused by highly pathogenic Porcine reproductive and respiratory syndrome virus.

    PubMed

    Cao, Jianbo; Li, Bin; Fang, Liurong; Chen, Huanchun; Xiao, Shaobo

    2012-07-01

    Since 2006, an unprecedented epidemic of highly pathogenic Porcine reproductive and respiratory syndrome virus (HP-PRRSV) infection has emerged and prevailed in mainland China, causing so called high fever disease with a nervous symptom that is different from typical porcine reproductive and respiratory syndrome. To investigate this syndrome, the brains of pigs inoculated with HP-PRRSV were analyzed. The nucleic acid of HP-PRRSV was detected in brains by reverse transcription polymerase chain reaction. Histological examination demonstrated nonsuppurative encephalitis with lymphohistiocytic perivascular cuffing and infiltration of these leukocytes into the neuropil. Furthermore, transmission electron microscopy revealed that the HP-PRRSV that infected the endothelial cells crossed the blood-brain barrier into the central nervous system then induced cellular damage to neurons and neuroglial cells. These results provide a general insight into the pathway of HP-PRRSV invasion into brain tissue and the pathogenesis of nonsuppurative encephalitis. PMID:22585954

  7. Production and Evaluation of Virus-Like Particles Displaying Immunogenic Epitopes of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV)

    PubMed Central

    Murthy, Ambika Mosale Venkatesh; Ni, Yanyan; Meng, Xiangjin; Zhang, Chenming

    2015-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is the most significant infectious disease currently affecting the swine industry worldwide. Several inactivated and modified live vaccines (MLV) have been developed to curb PRRSV infections. However, the efficacy and safety of these vaccines are unsatisfactory, and hence, there is a strong demand for the development of new PRRS universal vaccines. Virus-like particle (VLP)-based vaccines are gaining increasing acceptance compared to subunit vaccines, as they present the antigens in a more veritable conformation and are readily recognized by the immune system. Hepatitis B virus core antigen (HBcAg) has been successfully used as a carrier for more than 100 viral sequences. In this study, hybrid HBcAg VLPs were generated by fusion of the conserved protective epitopes of PRRSV and expressed in E. coli. An optimized purification protocol was developed to obtain hybrid HBcAg VLP protein from the inclusion bodies. This hybrid HBcAg VLP protein self-assembled to 23-nm VLPs that were shown to block virus infection of susceptible cells when tested on MARC 145 cells. Together with the safety of non-infectious and non-replicable VLPs and the low cost of production through E. coli fermentation, this hybrid VLP could be a promising vaccine candidate for PRRS. PMID:25874763

  8. Peptide nanofiber hydrogel adjuvanted live virus vaccine enhances cross-protective immunity to porcine reproductive and respiratory syndrome virus.

    PubMed

    Li, Xiangdong; Galliher-Beckley, Amy; Huang, Hongzhou; Sun, Xiuzhi; Shi, Jishu

    2013-09-23

    Porcine reproductive and respiratory syndrome virus (PRRSV) is prevalent in swine farms worldwide and is a major source of economic loss and animal suffering. Rapid genetic variation of PRRSV makes it difficult for current vaccines to confer protection against newly emerging strains. We recently demonstrated that a novel peptide nanofiber hydrogel (H9e) could act as a potent adjuvant for killed H1N1 vaccines. Therefore, the objective of this study was to evaluate H9e as an adjuvant for PRRSV modified live virus (MLV) vaccines. Pigs were vaccinated with Ingelvac PRRSV MLV with or without H9e adjuvant before being challenged with the VR-2332 (parental vaccine strain) or MN184A (genetically diverse strain) PRRSV. Pigs vaccinated with MLV+H9e had higher levels of circulating vaccine virus. More importantly, pigs vaccinated with MLV+H9e had improved protection against challenge by both PRRSV strains, as demonstrated by reduced challenge-induced viremia compared with pigs vaccinated with MLV alone. Pigs vaccinated with MLV+H9e had lower frequency of T-regulatory cells and IL-10 production but higher frequency of Th/memory cells and IFN-? secretion than that in pigs vaccinated with MLV alone. Taken together, our studies suggest that the peptide nanofiber hydrogel H9e, when combined with the PRRSV MLV vaccine, can enhance vaccine efficacy against two different PRRSV strains by modulating both host humoral and cellular immune responses. PMID:23933333

  9. Coincidence of Varicella-Zoster Virus Anterior Uveitis in a Patient with Chandler's Syndrome

    PubMed Central

    Joko, Takeshi; Suzuki, Takashi; Inoue, Tomoyuki; Kikuchi, Masaaki; Hara, Yuko; Shiraishi, Atsushi; Ohashi, Yuichi

    2013-01-01

    Purpose We report a patient who, based on the clinical manifestations, was originally diagnosed as having Chandler's syndrome and later developed varicella-zoster virus (VZV) DNA-positive anterior uveitis. Methods The patient with Chandler's syndrome who manifested anterior uveitis underwent a complete ophthalmologic examination. Polymerase chain reaction (PCR) was used to amplify the viral DNA in the aqueous humor to determine the cause of the intraocular inflammation. Results Slit-lamp biomicroscopy showed focal iris atrophy and peripheral anterior synechiae (PAS); specular microscopy of the corneal endothelium disclosed the hammered-silver appearance. Based on these clinical findings, we diagnosed this patient as having Chandler's syndrome. During the follow-up period, however, the inflammatory cells suddenly appeared in the anterior chamber with formation of keratic precipitates and an increased intraocular pressure (IOP). VZV DNA was displayed in the aqueous humor by PCR. Based upon the diagnosis of VZV anterior uveitis, corticosteroids and acyclovir were given topically and systemically. The inflammation subsided with these medications; however, trabeculectomy was finally needed to control the IOP due to PAS progression. Conclusion The coincidence of VZV anterior uveitis with Chandler's syndrome may constitute an implication for the possible viral etiology of iridocorneal endothelial syndrome. PMID:24474927

  10. Porcine reproductive and respiratory syndrome virus vaccine does not fit in classical vaccinology

    PubMed Central

    2015-01-01

    All vaccines are developed to elicit an effective immune response in vaccinated animals such as innate, humoral and cell mediated response to protect animal health. Quality and intensity of the immune responses are differing by characteristics of the vaccine formulation and nature of the infectious agent. Modified live virus vaccines showed advantages over killed vaccines in terms of rapid immune response, duration of the immunity and better cell mediated protection mechanism. The porcine reproductive and respiratory syndrome virus (PRRSV) is relatively newly emerging (1986 in United States, 1990 in Europe) viral pathogen in pigs and tremendous effort has been made to protect pigs from this economically devastating disease such as developing killed, modified live, recombinant protein based and DNA vaccines. However, only cell culture attenuated virus vaccine is practiced with arguably limited efficacy. The PRRSV vaccine did not clear virus from infected pigs nor prevent re-infection of the virus. The vaccine showed very limited innate immune response, low anamnestic immune response and negligible cell mediated immune response. Despite of the current developed scientific technology, there still remain many questions to solve a most important pig disease worldwide. PMID:26273574

  11. DNA condensates organized by the capsid protein VP15 in White Spot Syndrome Virus

    SciTech Connect

    Liu Yingjie; Wu Jinlu; Chen Hu; Hew, Choy Leong; Yan Jie

    2010-12-20

    The White Spot Syndrome Virus (WSSV) has a large circular double-stranded DNA genome of around 300 kb and it replicates in the nucleus of the host cells. The machinery of how the viral DNA is packaged has been remained unclear. VP15, a highly basic protein, is one of the major capsid proteins found in the virus. Previously, it was shown to be a DNA binding protein and was hypothesized to participate in the viral DNA packaging process. Using Atomic Force Microscopy imaging, we show that the viral DNA is associated with a (or more) capsid proteins. The organized viral DNA qualitatively resembles the conformations of VP15 induced DNA condensates in vitro. Furthermore, single-DNA manipulation experiments revealed that VP15 is able to condense single DNA against forces of a few pico Newtons. Our results suggest that VP15 may aid in the viral DNA packaging process by directly condensing DNA.

  12. The effects of a thermophile metabolite, tryptophol, upon protecting shrimp against white spot syndrome virus.

    PubMed

    Zhu, Fei; Jin, Min

    2015-12-01

    White spot syndrome virus (WSSV) is a shrimp pathogen responsible for significant economic loss in commercial shrimp farms and until now, there has been no effective approach to control this disease. In this study, tryptophol (indole-3-ethanol) was identified as a metabolite involved in bacteriophage-thermophile interactions. The dietary addition of tryptophol reduced the mortality in shrimp Marsupenaeus japonicus when orally challenged with WSSV. Our results revealed that 50 mg/kg tryptophol has a better protective effect in shrimp than 10 or 100 mg/kg tryptophol. WSSV copies in shrimp were reduced significantly (P < 0.01) when supplemented with 50 mg/kg tryptophol, indicating that virus replication was inhibited by tryptophol. Consequently, tryptophol represents an effective antiviral dietary supplement for shrimp, and thus holds significant promise as a novel and efficient therapeutic approach to control WSSV in shrimp aquaculture. PMID:26492993

  13. Mild Clinical Course of Severe Fever with Thrombocytopenia Syndrome Virus Infection in an Elderly Japanese Patient

    PubMed Central

    Ohagi, Yuko; Nakamoto, Chiaki; Nakamoto, Hiromichi; Saijo, Masayuki; Shimojima, Masayuki; Nakano, Yoshio; Fujimoto, Tokuzo

    2014-01-01

    Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious and hemorrhagic disease recently described in China and western Japan. A 71-year-old healthy Japanese woman noticed a tick biting her after harvesting in an orchard and removed it herself. She developed diarrhea, anorexia, and chills eight days later. Because these symptoms continued, she visited a primary care physician 6 days after the onset. Laboratory data revealed thrombocytopenia, leukocytopenia, and elevated liver enzymes. She was then referred to our hospital. Although not completely fulfilling the diagnostic criteria used in a retrospective study in Japan, SFTS was suspected, and we detected SFTS virus in the patient's blood using RT-PCR. However, she recovered without intensive treatment and severe complications 13 days after the onset. In this report, we present a mild clinical course of SFTS virus infection in Japan in detail. PMID:25574405

  14. Plant-based porcine reproductive and respiratory syndrome virus VLPs induce an immune response in mice.

    PubMed

    Uribe-Campero, Laura; Monroy-García, Alberto; Durán-Meza, Ana L; Villagrana-Escareño, María V; Ruíz-García, Jaime; Hernández, Jesús; Núñez-Palenius, Héctor G; Gómez-Lim, Miguel A

    2015-10-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) significantly affects the swine industry worldwide. An efficient, protective vaccine is still lacking. Here, we report for the first time the generation and purification of PRRSV virus like particles (VLPs) by expressing GP5, M and N genes in Nicotiana silvestris plants. The particles were clearly visible by transmission electron microscopy (TEM) with a size of 60-70 nm. Hydrodynamic diameter of the particles was obtained and it was confirmed that the VLPs had the appropriate size for PRRS virions and that the VLPs were highly pure. By measuring the Z potential we described the electrophoretic mobility behavior of VLPs and the best conditions for stability of the VLPs were determined. The particles were immunogenic in mice. A western blot of purified particles allowed detection of three coexpressed genes. These VLPs may serve as a platform to develop efficient PRRSV vaccines. PMID:26412521

  15. Cytokine production in immortalized porcine alveolar macrophages infected with porcine reproductive and respiratory syndrome virus.

    PubMed

    Lee, Yoo Jin; Lee, Changhee

    2012-12-15

    Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically significant viral pathogens for pig production worldwide. PRRSV primarily infects cells of the monocyte/macrophage lineage such as porcine alveolar macrophages (PAMs), and is generally known to suppress normal macrophage function and regulate innate immune responses to viral infection. A continuous PRRSV-permissive porcine monocyte-derived cell line was previously generated to facilitate virus propagation and advance research on the biology and immunology of PRRSV. With the availability of this valuable tool, we first sought to explore modulation of inflammatory cytokine expression in PAM-pCD163 cells infected with each genotype PRRSV and to establish an in vitro system for immune function studies using PRRSV isolates. PMID:23041033

  16. Vanishing bile duct syndrome in human immunodeficiency virus: Nevirapine hepatotoxicity revisited

    PubMed Central

    Kochar, Rajan; Nevah, Moises I; Lukens, Frank J; Fallon, Michael B; Machicao, Victor I

    2010-01-01

    Vanishing bile duct syndrome (VBDS) refers to a group of disorders characterized by prolonged cholestasis as a result of destruction and disappearance of intrahepatic bile ducts. Multiple etiologies have been indentified including infections, neoplastic disorders, autoimmune conditions and drugs. The natural history of this condition is variable and may involve resolution of cholestasis or progression with irreversible damage. VBDS is extremely rare in human immunodeficiency virus (HIV)-infected patients and anti-retroviral therapy has never been implicated as a cause. We encountered a young pregnant female with HIV and VBDS secondary to anti-retroviral therapy. Here, we report her clinical course and outcome. PMID:20614492

  17. Pathogenesis and prevention of placental and transplacental porcine reproductive and respiratory syndrome virus infection

    PubMed Central

    2013-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV)-induced reproductive problems are characterized by embryonic death, late-term abortions, early farrowing and increase in number of dead and mummified fetuses, and weak-born piglets. The virus recovery from fetal tissues illustrates transplacental infection, but despite many studies on the subject, the means by which PRRSV spreads from mother to fetus and the exact pathophysiological basis of the virus-induced reproductive failure remain unexplained. Recent findings from our group indicate that the endometrium and placenta are involved in the PRRSV passage from mother to fetus and that virus replication in the endometrial/placental tissues can be the actual reason for fetal death. The main purpose of this review is to clarify the role that PRRSV replication and PRRSV-induced changes in the endometrium/placenta play in the pathogenesis of PRRSV-induced reproductive failure in pregnant sows. In addition, strategies to control placental and transplacental PRRSV infection are discussed. PMID:24099529

  18. Inhibition of porcine reproductive and respiratory syndrome virus by Cecropin D in vitro.

    PubMed

    Liu, Xiaohong; Guo, Chunhe; Huang, Yumao; Zhang, Xiaoyu; Chen, Yaosheng

    2015-08-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause substantial economic losses to the pig industry worldwide. Although vaccines are commercially available for the control of PRRSV infection, no vaccination regimen has been proved sustained success in terms of generating a protective immune response. Therefore, the development of novel antivirals is urgently needed. Antimicrobial peptides display broad-spectrum antimicrobial activities against bacteria, fungi, and viruses and play an important role in host innate immune response. Here, we tested whether Cecropin D (CD) could inhibit PRRSV infection and replication in vitro. The inhibitory effect of CD occurred during viral attachment and the early period of viral entry into Marc-145 cells. CD also attenuated virus-induced apoptosis during the late phase of PRRSV infection and suppressed virus release in Marc-145 cells, which might contribute to the inhibition of PRRSV infection. Similar inhibitory effects on PRRSV infection were also found with CD treatment in porcine alveolar macrophages, the major target cell type of PRRSV infection in pigs in vivo. These findings suggest that CD has the potential to develop a new therapeutic agent against PRRSV infection. PMID:26102162

  19. Egg drop syndrome virus enters duck embryonic fibroblast cells via clathrin-mediated endocytosis.

    PubMed

    Huang, Jingjing; Tan, Dan; Wang, Yang; Liu, Caihong; Xu, Jiamin; Wang, Jingyu

    2015-12-01

    Previous studies of egg drop syndrome virus (EDSV) is restricted to serological surveys, disease diagnostics, and complete viral genome analysis. Consequently, the infection characteristics and entry routes of EDSV are poorly understood. Therefore, we aimed to explore the entry pathway of EDSV into duck embryonic fibroblast (DEF) cells as well as the infection characteristics and proliferation of EDSV in primary DEF and primary chicken embryo liver (CEL) cells. Transmission electron microscopy revealed that the virus triggered DEF cell membrane invagination as early as 10min post-infection and that integrated endocytic vesicles formed at 20min post-infection. The virus yield in EDSV-infected DEF cells treated with chlorpromazine (CPZ), sucrose, methyl-?-cyclodextrin (M?CD), or NH4Cl was measured by quantitative real-time PCR. Compared with the mock treatment, CPZ and sucrose greatly inhibited the production of viral progeny in a dose-dependent manner, while M?CD treatment did not result in a significant difference. Furthermore, NH4Cl had a strong inhibitory effect on the production of EDSV progeny. In addition, indirect immunofluorescence demonstrated that virus particles clustered on the surface of DEF cells treated with CPZ or sucrose. These results indicate that EDSV enters DEF cells through clathrin-mediated endocytosis followed by a pH-dependent step, which is similar to the mechanism of entry of human adenovirus types 2 and 5. PMID:26200954

  20. Haemorrhagic smolt syndrome (HSS) in Norway: pathology and associated virus-like particles.

    PubMed

    Nylund, A; Plarre, H; Hodneland, K; Devold, M; Aspehaug, V; Aarseth, M; Koren, C; Watanabe, K

    2003-03-17

    Atlantic salmon Salmo salar pre-smolt, smolt and post-smolt, with clinical signs of haemorrhagic smolt syndrome (HSS) have been found in several locations along the Norwegian coast (Rogaland to Troms). Affected fish had pale gills and bleeding at the fin bases, but seemed to be in good physical condition with no obvious weight loss. The internal organs and body cavity showed distinct bleedings. Petechiae were found on the gastrointestinal tract, swim bladder and peritoneum, visceral adipose tissue, heart and somatic musculature. The liver was bright yellow and sometimes mottled with petechiae and ecchymoses. Acitic fluid was found in the visceral cavity and fluid was also present in the pericardial cavity. Histological examination revealed haemorrhage in most organs. The glomeruli were degenerated and the renal tubules were filled with erythrocytes. The aims of this study were to describe the pathology and discover, if possible, the aetiology of the HSS. Tissues were collected for light and transmission electron microscopy (TEM), immunofluorescence (IFAT), reverse transcription (RT)-PCR diagnostics (screening for infectious salmon anaemia virus [ISAV], viral haemorrhagic septicaemia virus [VHSV], salmon pancreas disease virus [SPDV], sleeping disease virus [SDV] and infectious haematopoetic necrosis virus [IHNV]), and tissue homogenates (heart, liver, kidney and spleen) were sterile-filtered and inoculated into cell cultures. Homogenates made from several tissues were also injected intraperitoneally into salmon and rainbow trout Oncorhynchus mykiss. The diagnostic tests revealed no consistent findings of any pathogens, with the exception of TEM which showed 2 types of virus-like particles: Type I was 50 to 60 nm in diameter and Type II about 50 nm in diameter. These virus-like particles were found in salmon from all farms affected by HSS and screened by TEM. Several different cells, blood vessel endothelial cells, endocardial cells, heart myofibres, and leukocytes were associated with the 2 virus-like particles. The Type I particle seems to be an infectious pancreatic necrosis (IPN)-like virus, while (based on the number of target cells, particle morphology, budding and uptake into target cells) Type II particle could be a togavirus. PMID:12718466

  1. Comparative analysis of differentially expressed genes in normal and white spot syndrome virus infected Penaeus monodon

    PubMed Central

    Leu, Jiann-Horng; Chang, Chih-Chin; Wu, Jin-Lu; Hsu, Chun-Wei; Hirono, Ikuo; Aoki, Takashi; Juan, Hsueh-Fen; Lo, Chu-Fang; Kou, Guang-Hsiung; Huang, Hsuan-Cheng

    2007-01-01

    Background White spot syndrome (WSS) is a viral disease that affects most of the commercially important shrimps and causes serious economic losses to the shrimp farming industry worldwide. However, little information is available in terms of the molecular mechanisms of the host-virus interaction. In this study, we used an expressed sequence tag (EST) approach to observe global gene expression changes in white spot syndrome virus (WSSV)-infected postlarvae of Penaeus monodon. Results Sequencing of the complementary DNA clones of two libraries constructed from normal and WSSV-infected postlarvae produced a total of 15,981 high-quality ESTs. Of these ESTs, 46% were successfully matched against annotated genes in National Center of Biotechnology Information (NCBI) non-redundant (nr) database and 44% were functionally classified using the Gene Ontology (GO) scheme. Comparative EST analyses suggested that, in postlarval shrimp, WSSV infection strongly modulates the gene expression patterns in several organs or tissues, including the hepatopancreas, muscle, eyestalk and cuticle. Our data suggest that several basic cellular metabolic processes are likely to be affected, including oxidative phosphorylation, protein synthesis, the glycolytic pathway, and calcium ion balance. A group of immune-related chitin-binding protein genes is also likely to be strongly up regulated after WSSV infection. A database containing all the sequence data and analysis results is accessible at . Conclusion This study suggests that WSSV infection modulates expression of various kinds of genes. The predicted gene expression pattern changes not only reflect the possible responses of shrimp to the virus infection but also suggest how WSSV subverts cellular functions for virus multiplication. In addition, the ESTs reported in this study provide a rich source for identification of novel genes in shrimp. PMID:17506900

  2. Pharmacologic management of human immunodeficiency virus wasting syndrome.

    PubMed

    Badowski, Melissa; Pandit, Neha Sheth

    2014-08-01

    Pharmacologic interventions for human immunodeficiency virus (HIV) wasting have been studied since the 1990s, but the results of these interventions have been difficult to compare because the studies used different HIV wasting definitions and assessed various patient outcomes. Thus, we performed a systematic review of the current literature to identify studies that evaluated pharmacologic management of HIV wasting and to compare and contrast treatment options. Further, we provide a comprehensive review of these treatment options and describe the definition of HIV wasting used in each study, the outcomes assessed, and whether antiretroviral therapy was used during the HIV wasting treatment. Literature searches of the PubMed/Medline (1946-2014) and Google Scholar databases were performed, and a review of the bibliographies of retrieved articles was performed to identify additional references. Only English-language articles pertaining to humans and HIV-infected individuals were evaluated. Thirty-six studies were identified that assessed pharmacologic interventions to treat HIV wasting. Appetite stimulants, such as megestrol acetate, have been shown to increase total body weight (TBW) and body mass index in HIV-infected patients with wasting. Studies evaluating dronabinol showed conflicting data on TBW increases, but the drug may have minimal benefit on body composition compared with other appetite stimulants. Testosterone has been shown to be effective in HIV wasting for those who suffer from hypogonadism. Recombinant human growth hormone has been evaluated for HIV wasting and has shown promising results for TBW and lean body mass increases. Thalidomide has been studied; however, its use is limited due to its toxicities. Although megestrol acetate and dronabinol are approved by the U.S. Food and Drug Administration (FDA) for the treatment of HIV wasting, it is important to recognize other comorbidities such as depression or hypogonadism that may contribute to the patient's appetite and weight loss. If a patient is diagnosed with hypogonadism and HIV wasting, testosterone would be a good therapeutic option. Although mirtazapine is not FDA approved for the management of HIV wasting, it has been shown to promote weight gain while treating depression symptoms. Mirtazapine may be a promising pharmacologic option in the management of HIV wasting and depression, but further research is needed. PMID:24782295

  3. Chimeric viruses containing the N-terminal ectodomains of GP5 and M proteins of porcine reproductive and respiratory syndrome do not change the cellular tropism of equine arteritis virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Equine arteritis virus (EAV) and porcine reproductive and respiratory syndrome virus (PRRSV) are members of family Arteriviridae; they share many biological properties but differ significantly in cellular tropism. Using an infectious cDNA clone of EAV, we engineered a panel of six chimeric viruses b...

  4. Screening, isolation and optimization of anti–white spot syndrome virus drug derived from terrestrial plants

    PubMed Central

    Ghosh, Upasana; Chakraborty, Somnath; Balasubramanian, Thangavel; Das, Punyabrata

    2014-01-01

    Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various terrestrial plants and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti–WSSV property in Litopenaeus vannamei. The best anti–WSSV plant isolate, TP22C was isolated and further analyzed. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug. Results Seven plant isolates exhibited significant survivability in host. The drug TP22C thus formulated showed 86% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of TP22C required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 750 mg/kg body weight/day survived at the rate of 86%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug TP22C derived from Momordica charantia is a potent anti-white spot syndrome virus drug. PMID:25183066

  5. Pathogenicity and molecular characterization of emerging porcine reproductive and respiratory syndrome virus in Vietnam in 2007.

    PubMed

    Metwally, S; Mohamed, F; Faaberg, K; Burrage, T; Prarat, M; Moran, K; Bracht, A; Mayr, G; Berninger, M; Koster, L; To, T L; Nguyen, V L; Reising, M; Landgraf, J; Cox, L; Lubroth, J; Carrillo, C

    2010-10-01

    In 2007, Vietnam experienced swine disease outbreaks causing clinical signs similar to the 'porcine high fever disease' that occurred in China during 2006. Analysis of diagnostic samples from the disease outbreaks in Vietnam identified porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV-2). Additionally, Escherichia coli and Streptococcus equi subspecies zooepidemicus were cultured from lung and spleen, and Streptococcus suis from one spleen sample. Genetic characterization of the Vietnamese PRRSV isolates revealed that this virus belongs to the North American genotype (type 2) with a high nucleotide identity to the recently reported Chinese strains. Amino acid sequence in the nsp2 region revealed 95.7-99.4% identity to Chinese strain HUN4, 68-69% identity to strain VR-2332 and 58-59% identity to strain MN184. A partial deletion in the nsp2 gene was detected; however, this deletion did not appear to enhance the virus pathogenicity in the inoculated pigs. Animal inoculation studies were conducted to determine the pathogenicity of PRRSV and to identify other possible agents present in the original specimens. Pigs inoculated with PRRSV alone and their contacts showed persistent fever, and two of five pigs developed cough, neurological signs and swollen joints. Necropsy examination showed mild to moderate bronchopneumonia, enlarged lymph nodes, fibrinous pericarditis and polyarthritis. PRRSV was re-isolated from blood and tissues of the inoculated and contact pigs. Pigs inoculated with lung and spleen tissue homogenates from sick pigs from Vietnam developed high fever, septicaemia, and died acutely within 72 h, while their contact pigs showed no clinical signs throughout the experiment. Streptococcus equi subspecies zooepidemicus was cultured, and PRRSV was re-isolated only from the inoculated pigs. Results suggest that the cause of the swine deaths in Vietnam is a multifactorial syndrome with PRRSV as a major factor. PMID:20629970

  6. Risks and prevention of severe RS virus infection among children with immunodeficiency and Down's syndrome.

    PubMed

    Mori, Masaaki; Morio, Tomohiro; Ito, Shuichi; Morimoto, Akira; Ota, Setsuo; Mizuta, Koichi; Iwata, Tsutomu; Hara, Toshiro; Saji, Tsutomu

    2014-08-01

    By the age of two years, almost all infants are infected with the Respiratory syncytial virus (RSV). One of the main causes of hospitalizations for bronchiolitis and pneumonia at this age is RSV infection. In addition to well-known risks for severe RSV disease, such as prematurity, bronchopulmonary dysplasia and congenital heart disease, immunodeficiencies, chromosomal abnormalities such as Down's syndrome or neuromuscular diseases have also been identified as risks. While the medical needs for RSV prevention in these risk groups are high, clinical evidence to support this is limited. Palivizumab was recently approved in Japan for prophylaxis in children with immunodeficiency or Down's syndrome. A clinical guidance protocol for the prevention of RSV infection using Palivizumab in these risk groups is provided here on the basis of a review of the available literature and on expert opinion. Thus, the present article reviews the published literature related to RSV infections in infants and children with immunodeficiencies or Down's syndrome in order to outline the risks, pathology and physiology of severe RSV disease in these patient groups. The purpose of this article is to facilitate understanding of the medical scientific bases for the clinical guidance. PMID:24929631

  7. Haemaphysalis longicornis Ticks as Reservoir and Vector of Severe Fever with Thrombocytopenia Syndrome Virus in China

    PubMed Central

    Luo, Li-Mei; Zhao, Li; Wen, Hong-Ling; Zhang, Zhen-Tang; Liu, Jian-Wei; Fang, Li-Zhu; Xue, Zai-Feng; Ma, Dong-Qiang; Zhang, Xiao-Shuang; Ding, Shu-Jun; Lei, Xiao-Ying

    2015-01-01

    Severe fever with thrombocytopenia syndrome (SFTS) is an emerging hemorrhagic fever in East Asia caused by SFTS virus (SFTSV), a newly discovered phlebovirus. The Haemaphysalis longicornis tick has been suspected to be the vector of SFTSV. To determine whether SFTSV can be transmitted among ticks, from ticks to animals, and from animals to ticks, we conducted transmission studies between developmental stages of H. longicornis ticks and between ticks and mice. Using reverse transcription PCR, we also analyzed the prevalence of SFTSV infection among H. longicornis ticks collected from vegetation in Shandong Province, China. Our results showed a low prevalence of SFTSV among collected ticks (0.2%, 8/3,300 ticks), and we showed that ticks fed on SFTSV-infected mice could acquire the virus and transstadially and transovarially transmit it to other developmental stages of ticks. Furthermore, SFTSV-infected ticks could transmit the virus to mice during feeding. Our findings indicate ticks could serve as a vector and reservoir of SFTSV. PMID:26402039

  8. N-glycosylation profiling of porcine reproductive and respiratory syndrome virus envelope glycoprotein 5.

    PubMed

    Li, Juan; Tao, Shujuan; Orlando, Ron; Murtaugh, Michael P

    2015-04-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a positive-sense ssRNA virus whose envelope contains four glycoproteins and three nonglycosylated proteins. Glycans of major envelope glycoprotein 5 (GP5) are proposed as important for virus assembly and entry into permissive cells. Structural characterization of GP5 glycans would facilitate the mechanistic understanding of these processes. Thus, we purified the PRRSV type 2 prototype strain, VR2332, and analyzed the virion-associated glycans by both biochemical and mass spectrometric methods. Endoglycosidase digestion showed that GP5 was the primary protein substrate, and that the carbohydrate moieties were primarily complex-type N-glycans. Mass spectrometric analysis (HPLC-ESI-MS/MS) of GP5 N-glycans revealed an abundance of N-acetylglucosamine (GlcNAc) and N-acetyllactosamine (LacNAc) oligomers in addition to sialic acids. GlcNAc and LacNAc accessibility to ligands was confirmed by lectin co-precipitation. Our findings help to explain PRRSV infection of cells lacking sialoadhesin and provide a glycan database to facilitate molecular structural studies of PRRSV. PMID:25726973

  9. Function of CD163 fragments in porcine reproductive and respiratory syndrome virus infection

    PubMed Central

    Li, Hong; Zhou, En-Min; Liu, Cheng-Qian; Yi, Jian-Zhong

    2015-01-01

    Monocyte/macrophage scavenger receptor CD163 plays an important role in porcine reproductive and respiratory syndrome virus (PRRSV) infection. To identify the domains of CD163 involved in PRRSV infection, CD163 fragments P1 (1-798 bp), P2 (790-2046 bp), P3 (2023-3345 bp), Y-P1 (160-798 bp), Y-P2 (790-2046 bp) and Y-P3 (2143-3084 bp) were expressed by eukaryotic and prokaryotic expression systems, respectively. Infection experiments revealed that non-permissive BHK-21 cells transfected with pCD163 could be infected by PRRSV. However, cells with truncated CD163 (P1, P2, or P3) were not susceptible to PRRSV. Meanwhile, Y-P1, Y-P2, and Y-P3 were expressed in E. coli and antisera to these peptides were prepared in mice. A virus blocking test showed that Y-P2 protein and anti-Y-P2 mouse serum could block PRRSV infection in a dose-dependent manner, while Y-P3 protein could improve virus infection. PMID:26629026

  10. Pathogenicity of swine influenza viruses possessing an avian or swine-origin PB2 polymerase gene evaluated in mouse and pig models

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Influenza A viruses isolated from birds normally contain a PB2 polymerase gene with the avian-signature glutamic acid (E) at position 627, while those isolated from humans contain the mammalian-signature lysine (K) at this position. This residue has been shown to be a determinant of host range and c...

  11. Clinical and pathological responses of pigs from two genetically diverse commercial lines to porcine respiratory and reproductive syndrome virus infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The response to infection from porcine reproductive and respiratory syndrome virus (PRRSV) for two genetically diverse commercial pig lines was investigated. Seventy two pigs from each line, aged 6 weeks, were challenged with PRRSV VR-2385, and 66 littermates served as control. The clinical response...

  12. Differential immunity in pigs with high and low responses to porcine reproductive and respiratory syndrome virus (PRRSV) infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One hundred Hampshire by Duroc crossbred pigs (HD) and 100 NE Index line pigs (I) were infected with porcine reproductive and respiratory syndrome virus (PRRSV) and evaluated for resistance/susceptibility. Controls (100/line) were uninfected littermates to infected pigs. Viremia (V), weight change (...

  13. Whole blood microarray analysis of pigs showing extreme phenotypes after a porcine reproductive and respiratory syndrome virus infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background Observed variability in pig response to Porcine Reproductive and Respiratory Syndrome virus (PRRSv) infection, and recently demonstrated genetic control of such responses, suggest that it may be possible to reduce the economic impact of this disease by selecting more disease-resistant pig...

  14. Health Administrator Perspectives on Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome Prevention and Services at Historically Black Colleges and Universities

    ERIC Educational Resources Information Center

    Warren-Jeanpiere, Lari; Jones, Sandra; Sutton, Madeline Y.

    2011-01-01

    Objective: Due to the disproportionate impact of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) among African American young adults, the authors explored (1) number of historically black college and university (HBCU) campuses with existing HIV prevention policies and services and (2) perceived barriers for implementing…

  15. Pathogenicity of three type 2 Porcine Reproductive and Respiratory Syndrome virus strains in experimentally inoculated pregnant gilts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mechanisms of reproductive failure resulting from infection with porcine reproductive and respiratory syndrome virus (PRRSv) are still poorly understood. The present study, a side-by-side evaluation of the pathogenicity of three type 2 PRRSv strains in a reproductive model, was used as a pilot study...

  16. Pathogenicity and distribution of highly pathogenic porcine reproductive and respiratory syndrome virus in pigs.

    PubMed

    Hu, S P; Zhang, Z; Liu, Y G; Tian, Z J; Wu, D L; Cai, X H; He, X J

    2013-08-01

    The pathogenesis of highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) strain (HuN4) is poorly understood. Therefore, highly pathogenic PRRSV strain (HuN4) and its derivative strain (HuN4-F112) (obtained by propagation in MARC145 cells for 112 passages) were inoculated into a total of 48 PRRSV-sero-negative pigs (age: 4-5 weeks) by the intranasal route. Virological, pathological and in situ hybridization analyses were performed. The results exhibited that pigs infected with HuN4 showed a loss of appetite, decrease in body weight, raised body temperature and respiratory symptoms, along with interstitial pneumonia lesions. In the HuN4 group, multifocal interstitial pneumonia with macrophage infiltration was found in the lung. The lesions in the lymph node were characterized by collapsed follicles, depletion of germinal centres and reduction in lymphocytes. Perivascular cuffing and glial nodules were observed in the brains of some pigs. By comparison, the HuN4-F112 group had milder lesions. PRRSV was detected in macrophages, alveolar epithelial cells and vascular endothelial cells in the tonsil and lymph nodes. The PRRSV amounts in the pigs infected with HuN4 were 10(5) -10(9) copies/ml in the blood and 10(10) -10(11) copies/g in the lung tissues, whereas the virus amounts with HuN4-F112 were 10(2.15) -10(3.13) copies/ml in the blood and 10(3.0) -10(3.6) copies/g in the lung. Our results demonstrate that the PRRS HuN4 virus infects alveolar epithelial cells, macrophages and vascular endothelial cells causing diffuse alveolar damage and lymph node necrosis. Its higher pathogenicity compared with HuN4-F112 virus may be explained in part by higher replication rate in the previously mentioned organs. PMID:22762447

  17. Crassostrea gigas oysters as a shrimp farm bioindicator of white spot syndrome virus.

    PubMed

    Vazquez-Boucard, C; Escobedo-Fregoso, C; Duran-Avelar, Ma de J; Mercier, L; Llera-Herrera, R; Escobedo-Bonilla, C; Vibanco-Perez, N

    2012-04-26

    This study explored whether Crassostrea gigas oysters can be used as a bioindicator of white spot syndrome virus (WSSV) in shrimp farm water canals. Bioassays showed that C. gigas can accumulate WSSV in their gills and digestive glands but do not become infected, either by exposure to seawater containing WSSV or by cohabitation with infected shrimp. The use of a WSSV nested PCR to screen oysters placed in water canals at the entry of a shrimp farm allowed WSSV to be detected 16 d prior to the disease occurring. The finding that C. gigas can concentrate small amounts of WSSV present in seawater without being harmed makes it an ideal sentinel species at shrimp farms. PMID:22535870

  18. Endocytic pathway is indicated for white spot syndrome virus (WSSV) entry in shrimp.

    PubMed

    Huang, Zih-Jhan; Kang, Shih-Ting; Leu, Jiann-Horng; Chen, Li-Li

    2013-09-01

    The white spot syndrome virus (WSSV) has had a serious economic impact on the global shrimp aquaculture industry in the past two decades. Although research has clarified a lot about its genome and structure, the mechanism of how WSSV enters a cell is still unclear. In this study to determine this mechanism, primary cultured hemocytes were used as an experimental model to observe the process of WSSV entry because the stable shrimp cell lines for WSSV infection are lacking. After labeling virions and endosomes with fluorescent dyes followed by observation with a confocal microscope, the results show that the WSSV colocalizes with early endosomes. Hemocytes are further treated with different endocytic inhibitors, methyl-?-cyclodextrin (M?CD) and chlorpromazine (CPZ). WSSV still can be detected in the hemocytes treated with CPZ, but not in the hemocytes treated with M?CD. Thus, we conclude that WSSV adopts the caveolae-mediated endocytosis to enter the shrimp cell. PMID:23747417

  19. Functional identification of the non-specific nuclease from white spot syndrome virus

    SciTech Connect

    Li Li; Lin Shumei; Yanga Feng . E-mail: mbiotech@public.xm.fj.cn

    2005-07-05

    The product encoded by the wsv191 gene from shrimp white spot syndrome virus (WSSV) is homologous with non-specific nucleases (NSN) of other organisms. To functionally identify the protein, the wsv191 gene was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein with 6His-tag at C-terminal. The fusion protein (termed as rWSSV-NSN) was purified using Ni-NTA affinity chromatography under denatured conditions, renatured and characterized by three methods. The results showed that rWSSV-NSN could hydrolyze both DNA and RNA. 5'-RACE result revealed that the transcription initiation site of the wsv191 gene was located at nucleotide residue G of the predicted ATG triplet. Therefore, we concluded that the next ATG should be the genuine translation initiation codon of the wsv191 gene. Western blot analysis revealed that the molecular mass of natural WSSV-NSN was 37 kDa.

  20. Expression, purification and crystallization of two major envelope proteins from white spot syndrome virus

    SciTech Connect

    Tang, Xuhua; Hew, Choy Leong

    2007-07-01

    The crystallization of the N-terminal transmembrane region-truncated VP26 and VP28 of white spot syndrome virus is described. White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 Å resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 Å. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 Å, and diffracts to 2.0 Å resolution.

  1. Cytokine profiles in pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus and relationships with viral load and fetal outcome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In spite of extensive research, immunologic control mechanisms against Porcine Reproductive and Respiratory Syndrome virus (PRRSv) remain poorly understood. Cytokine responses have been exhaustively studied in nursery pigs and show contradictory results. Since no detailed reports on cytokine respons...

  2. Transmission of white spot syndrome virus (WSSV) from Dendronereis spp. (Peters) (Nereididae) to penaeid shrimp.

    PubMed

    Haryadi, D; Verreth, J A J; Verdegem, M C J; Vlak, J M

    2015-05-01

    Dendronereis spp. (Peters) (Nereididae) is a common polychaete in shrimp ponds built on intertidal land and is natural food for shrimp in traditionally managed ponds in Indonesia. White spot syndrome virus (WSSV), an important viral pathogen of the shrimp, can replicate in this polychaete (Desrina et al. 2013); therefore, it is a potential propagative vector for virus transmission. The major aim of this study was to determine whether WSSV can be transmitted from naturally infected Dendronereis spp. to specific pathogen-free (SPF) Pacific white shrimp Litopenaeus vannamei (Boone) through feeding. WSSV was detected in naturally infected Dendronereis spp. and Penaeus monodon Fabricius from a traditional shrimp pond, and the positive animals were used in the current experiment. WSSV-infected Dendronereis spp. and P. monodon in a pond had a point prevalence of 90% and 80%, respectively, as measured by PCR. WSSV was detected in the head, gills, blood and mid-body of Dendronereis spp. WSSV from naturally infected Dendronereis spp was transmitted to SPF L. vannamei and subsequently from this shrimp to new naïve-SPF L. vannamei to cause transient infection. Our findings support the contention that Dendronereis spp, upon feeding, can be a source of WSSV infection of shrimp in ponds. PMID:24716813

  3. DRACO inhibits porcine reproductive and respiratory syndrome virus replication in vitro.

    PubMed

    Guo, Chunhe; Chen, Luxi; Mo, Delin; Chen, Yaosheng; Liu, Xiaohong

    2015-05-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause substantial economic losses to the pig industry worldwide. Current vaccination strategies and antiviral drugs against PRRSV are still inadequate. Therefore, there is an urgent need for new antiviral strategies to control PRRSV. Double-stranded RNA (dsRNA) Activated Caspase Oligomerizer (DRACO) is a synthetic construct consisting of a dsRNA detection domain, an apoptosis induction domain, and a transduction tag. It has been shown to have broad-spectrum antiviral activity, but there have been no reports regarding its effect on PRRSV. Here, we demonstrate that DRACO exhibits robust antiviral activity against PRRSV infection by suppressing virus RNA and protein synthesis in both Marc-145 cells and porcine alveolar macrophages (PAMs). In addition, DRACO still exhibited strong anti-PRRSV activity when viral replication was enhanced by knockdown of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) in Marc-145 cells. Furthermore, in PAMs, DRACO was capable of inducing IL-6 expression and reducing Hsp70 expression, which might contribute to the inhibition of PRRSV infection. Collectively, our results imply that DRACO holds promise as a novel anti-PRRSV therapeutic drug. PMID:25772577

  4. Human immunodeficiency virus/acquired immune deficiency syndrome: Using drug from mathematical perceptive.

    PubMed

    Chatterjee, Amar Nath; Saha, Shubhankar; Roy, Priti Kumar

    2015-11-12

    Entry of acquired immune deficiency syndrome virus into the host immune cell involves the participation of various components of host and viral cell unit. These components may be categorized as attachment of the viral surface envelope protein subunit, gp120, to the CD4(+) receptor and chemokine coreceptors, CCR5 and CXCR4, present on T cell surface. The viral fusion protein, gp41, the second cleaved subunit of Env undergoes reconfiguration and the membrane fusion reaction itself. Since the CD4(+) T cell population is actively involved; the ultimate outcome of human immunodeficiency virus infection is total collapse of the host immune system. Mathematical modeling of the stages in viral membrane protein-host cell receptor-coreceptor interaction and the effect of antibody vaccine on the viral entry into the susceptible host cell has been carried out using as impulsive differential equations. We have studied the effect of antibody vaccination and determined analytically the threshold value of drug dosage and dosing interval for optimum levels of infection. We have also investigated the effect of perfect adherence of drug dose on the immune cell count in extreme cases and observed that systematic drug dosage of the immune cells leads to longer and improved lives. PMID:26568917

  5. Human immunodeficiency virus/acquired immune deficiency syndrome: Using drug from mathematical perceptive

    PubMed Central

    Chatterjee, Amar Nath; Saha, Shubhankar; Roy, Priti Kumar

    2015-01-01

    Entry of acquired immune deficiency syndrome virus into the host immune cell involves the participation of various components of host and viral cell unit. These components may be categorized as attachment of the viral surface envelope protein subunit, gp120, to the CD4+ receptor and chemokine coreceptors, CCR5 and CXCR4, present on T cell surface. The viral fusion protein, gp41, the second cleaved subunit of Env undergoes reconfiguration and the membrane fusion reaction itself. Since the CD4+ T cell population is actively involved; the ultimate outcome of human immunodeficiency virus infection is total collapse of the host immune system. Mathematical modeling of the stages in viral membrane protein-host cell receptor-coreceptor interaction and the effect of antibody vaccine on the viral entry into the susceptible host cell has been carried out using as impulsive differential equations. We have studied the effect of antibody vaccination and determined analytically the threshold value of drug dosage and dosing interval for optimum levels of infection. We have also investigated the effect of perfect adherence of drug dose on the immune cell count in extreme cases and observed that systematic drug dosage of the immune cells leads to longer and improved lives. PMID:26568917

  6. Birth Weight, Intrauterine Growth Retardation and Fetal Susceptibility to Porcine Reproductive and Respiratory Syndrome Virus

    PubMed Central

    Ladinig, Andrea; Foxcroft, George; Ashley, Carolyn; Lunney, Joan K.; Plastow, Graham; Harding, John C. S.

    2014-01-01

    The severity of porcine reproductive and respiratory syndrome was compared in pregnant gilts originating from high and low birth weight litters. One-hundred and eleven pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus on gestation day 85 (±1) were necropsied along with their fetuses 21 days later. Ovulation rates and litter size did not differ between groups, but fetuses from low birth weight gilts were shorter, lighter and demonstrated evidence of asymmetric growth with large brain:organ weight ratios (i.e. brain sparing). The number of intrauterine growth retarded fetuses, defined by brain:organ weight ratios greater than 1 standard deviation from the mean, was significantly greater in low, compared to high, birth weight gilts. Although ?? T cells significantly decreased over time in high compared to low birth weight gilts, viral load in serum and tissues, gilt serum cytokine levels, and litter outcome, including the percent dead fetuses per litter, did not differ by birth weight group. Thus, this study provided no substantive evidence that the severity of porcine reproductive and respiratory syndrome is affected by dam birth weight. However, intrauterine growth retarded fetuses had lower viral loads in both fetal thymus and in endometrium adjacent to the umbilical stump. Crown rump length did not significantly differ between fetuses that survived and those that died at least one week prior to termination. Taken together, this study clearly demonstrates that birth weight is a transgenerational trait in pigs, and provides evidence that larger fetuses are more susceptible to transplacental PRRSv infection. PMID:25275491

  7. Innate and adaptive immunity against Porcine Reproductive and Respiratory Syndrome Virus.

    PubMed

    Loving, Crystal L; Osorio, Fernando A; Murtaugh, Michael P; Zuckermann, Federico A

    2015-09-15

    Many highly effective vaccines have been produced against viruses whose virulent infection elicits strong and durable protective immunity. In these cases, characterization of immune effector mechanisms and identification of protective epitopes/immunogens has been informative for the development of successful vaccine programs. Diseases in which the immune system does not rapidly clear the acute infection and/or convalescent immunity does not provide highly effective protection against secondary challenge pose a major hurdle for clinicians and scientists. Porcine reproductive and respiratory syndrome virus (PRRSV) falls primarily into this category, though not entirely. PRRSV causes a prolonged infection, though the host eventually clears the virus. Neutralizing antibodies can provide passive protection when present prior to challenge, though infection can be controlled in the absence of detectable neutralizing antibodies. In addition, primed pigs (through natural exposure or vaccination with a modified-live vaccine) show some protection against secondary challenge. While peripheral PRRSV-specific T cell responses have been examined, their direct contribution to antibody-mediated immunity and viral clearance have not been fully elucidated. The innate immune response following PRRSV infection, particularly the antiviral type I interferon response, is meager, but when provided exogenously, IFN-? enhances PRRSV immunity and viral control. Overall, the quality of immunity induced by natural PRRSV infection is not ideal for informing vaccine development programs. The epitopes necessary for protection may be identified through natural exposure or modified-live vaccines and subsequently applied to vaccine delivery platforms to accelerate induction of protective immunity following vaccination. Collectively, further work to identify protective B and T cell epitopes and mechanisms by which PRRSV eludes innate immunity will enhance our ability to develop more effective methods to control and eliminate PRRS disease. PMID:26209116

  8. Middle East respiratory syndrome coronavirus infection: virus-host cell interactions and implications on pathogenesis.

    PubMed

    Zhou, Jie; Chu, Hin; Chan, Jasper Fuk-Woo; Yuen, Kwok-Yung

    2015-01-01

    Middle-East Respiratory Syndrome coronavirus (MERS-CoV) was identified to cause severe respiratory infection in humans since 2012. The continuing MERS epidemic with a case-fatality of more than 30 % poses a major threat to public health worldwide. Currently, the pathogenesis of human MERS-CoV infection remains poorly understood. We reviewed experimental findings from human primary cells and ex vivo human lung tissues, as well as those from animal studies, so as to understand the pathogenesis and high case-fatality of MERS. Human respiratory epithelial cells are highly susceptible to MERS-CoV and can support productive viral replication. However, the induction of antiviral cytokines and proinflammatory cytokines/chemokines are substantially dampened in the infected epithelial cells, due to the antagonistic mechanisms evolved by the virus. MERS-CoV can readily infect and robustly replicate in human macrophages and dendritic cells, triggering the aberrant production of proinflammatory cytokines/chemokines. MERS-CoV can also effectively infect human primary T cells and induce massive apoptosis in these cells. Although data from clinical, in vitro and ex vivo studies suggested the potential for virus dissemination, extrapulmonary involvement in MERS patients has not been ascertained due to the lack of autopsy study. In MERS-CoV permissive animal models, although viral RNA can be detected from multiple organs of the affected animals, the brain of human DPP4-transgenic mouse was the only extrapulmonary organ from which the infectious virus can be recovered. More research findings on the pathogenesis of MERS and the tissue tropisms of MERS-CoV may help to improve the treatment and infection control of MERS. PMID:26690369

  9. Porcine Reproductive and Respiratory Syndrome Virus Comparison: Divergent Evolution on Two Continents

    PubMed Central

    Nelsen, Chris J.; Murtaugh, Michael P.; Faaberg, Kay S.

    1999-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a recently described arterivirus responsible for disease in swine worldwide. Comparative sequence analysis of 3?-terminal structural genes of the single-stranded RNA viral genome revealed the presence of two genotypic classes of PRRSV, represented by the prototype North American and European strains, VR-2332 and Lelystad virus (LV), respectively. To better understand the evolution and pathogenicity of PRRSV, we obtained the 12,066-base 5?-terminal nucleotide sequence of VR-2332, encoding the viral replication activities, and compared it to those of LV and other arteriviruses. VR-2332 and LV differ markedly in the 5? leader and sections of the open reading frame (ORF) 1a region. The ORF 1b sequence was nearly colinear but varied in similarity of proteins encoded in identified regions. Furthermore, molecular and biochemical analysis of subgenomic mRNA (sgmRNA) processing revealed extensive variation in the number of sgmRNAs which may be generated during infection and in the lengths of noncoding sequence between leader-body junctions and the translation-initiating codon AUG. In addition, VR-2332 and LV select different leader-body junction sites from a pool of similar candidate sites to produce sgmRNA 7, encoding the viral nucleocapsid protein. The presence of substantial variations across the entire genome and in sgmRNA processing indicates that PRRSV has evolved independently on separate continents. The near-simultaneous global emergence of a new swine disease caused by divergently evolved viruses suggests that changes in swine husbandry and management may have contributed to the emergence of PRRS. PMID:9847330

  10. The presence of alpha interferon at the time of infection alters the innate and adaptive immune responses to porcine reproductive and respiratory syndrome virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome (PRRS) is one of the most devastating and costly diseases to the swine industry world-wide. Overall, the adaptive immune response to PRRS virus (PRRSV) is weak and results in delayed elimination of virus from the host and inferior vaccine protection. PRR...

  11. Immunodominant epitopes in nsp2 of porcine reproductive and respiratory syndrome virus are dispensable for replication but play an important role in viral pathogenesis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The nonstructural protein 2 (nsp2) of porcine reproductive and respiratory syndrome virus (PRRSV) is the largest protein of the virus. Besides its crucial role in viral replication, recent studies indicated its involvement in modulating host immunity. In this study, each of the six identified immu...

  12. DETECTION OF PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS IN SEMEN AND SERUM OF BOARS DURING THE FIRST SIX DAYS AFTER INOCULATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome (PRRS) is the number one disease problem faced by pork producers in the US. It is caused by the PRRS virus (PRRSV) and this virus can be shed in the semen of infected boars. Transmission of PRRSV by way of contaminated semen is a significant problem fo...

  13. Immune responses of pigs immunized with a recombinant porcine reproductive and respiratory syndrome virus expressing porcine GM-CSF.

    PubMed

    Li, Zhijun; Wang, Gang; Wang, Yan; Zhang, Chong; Huang, Baicheng; Li, Qiongyi; Li, Liangliang; Xue, Biyun; Ding, Peiyang; Cai, Xuehui; Wang, Chengbao; Zhou, En-Min

    2015-11-15

    Porcine reproductive and respiratory syndrome virus (PRRSV) has spread worldwide, causing huge economic losses to the swine industry. The current PRRSV vaccines have failed to provide broad protection against various strains. Granulocyte macrophage colony-stimulating factor (GM-CSF), an efficacious adjuvant, has been shown to enhance the immunogenicity of various vaccines. The purpose of this study was to construct a recombinant live attenuated PRRSV that expresses porcine GM-CSF (pGM-CSF) and evaluate the immune responses of pigs immunized with the recombinant virus. The results showed that the recombinant PRRSV was successfully rescued and had similar growth properties to parental virus grown in Marc-145 cells. The recombinant virus was stable for 10 passages in cell culture. Pigs intramuscularly immunized with the recombinant virus produced a similar humoral response to that elicited using parental virus. With regard to cell-mediated immunity assessed in peripheral blood, the recombinant virus induced higher proportion of CD4(+)CD8(+) double-positive T cells (DPT), higher IFN-? level at 0 and 7 days post-challenge (DPC), and lower viremia at 21 DPC than pigs immunized with parental virus. These results indicate that recombinant PRRSV expressing pGM-CSF can induce a significant higher cellular immune response and reduce the persistent infection compared pigs vaccinated with the parental virus. This is first report of evaluation of immune response in pigs elicited by a recombinant live attenuated PRRSV expressing porcine GM-CSF. It may represent a novel strategy for future development of genetic engineered vaccines against PRRSV infection. PMID:26300317

  14. Torque teno sus virus 1 and 2 viral loads in postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS) affected pigs.

    PubMed

    Aramouni, M; Segalés, J; Sibila, M; Martin-Valls, G E; Nieto, D; Kekarainen, T

    2011-12-15

    Torque teno viruses (TTV) are small, non-enveloped viruses with a circular single-stranded DNA genome, which are considered non-pathogenic. However, TTVs have been eventually linked to human diseases. TTVs infecting pigs, Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2), have been recently associated to porcine circovirus diseases (PCVD). To get more insights into such potential disease association, the aim of this study was to quantify TTSuV1 and TTSuV2 viral loads in serum of pigs affected by two PCVDs, postweaning multisystemic wasting syndrome (PMWS) and porcine dermatitis and nephropathy syndrome (PDNS). Such study was carried out by means of a newly developed real-time quantitative PCR (qPCR) method. Both TTSuVs were highly prevalent among studied pigs. TTSuV2 viral loads were significantly higher in PMWS affected animals, further supporting the previously suggested association between TTSuV2 and PMWS. On the contrary, TTSuV1 prevalence and loads were not related with the studied PCVDs. PMID:21719215

  15. Superior Orbital Fissure Syndrome and Ophthalmoplegia Caused by Varicella Zoster Virus with No Skin Eruption in a Patient Treated with Tumor Necrosis Alpha Inhibitor

    PubMed Central

    Jensen, Helene; Thomsen, Sidsel Thorup; Hansen, Stine Scott; Munksgaard, Signe Bruun; Lindelof, Mette

    2015-01-01

    Varicella zoster virus lies dormant in the dorsal root ganglia after symptomatic chicken pox infection, usually in childhood. If the virus reactivates in the trigeminal ganglia, it can cause varicella zoster ophthalmicus, which can have severe ocular complications. We report a case of a 73-year-old woman in severe immunosuppression due to treatment with mycophenolate mofetil, glucocorticosteroids and a tumor necrosis factor alpha inhibitor. The reactivation caused superior orbital fissure syndrome, which has only rarely been described in relation to varicella zoster virus reactivation. In our case, the syndrome was seen along with severe encephalitis. PMID:26600786

  16. Retroviral sequences related to human T-lymphotropic virus type II in patients with chronic fatigue immune dysfunction syndrome

    SciTech Connect

    DeFreitas, E.; Hilliard, B.; Cheney, P.R.; Bell, D.S.; Kiggundu, E.; Sankey, D.; Wroblewska, Z.; Palladino, M.; Woodward, J.P.; Koprowski, H. )

    1991-04-01

    Chronic fatigue immune dysfunction syndrome (CFIDS) is a recently recognized illness characterized by debilitating fatigue as well as immunological and neurological abnormalities. Once thought to be caused by Epstein-Barr virus, it is now thought to have a different but unknown etiology. The authors evaluted 30 adult and pediatric CFIDS patients from six eastern states for the presence of human T-lymphotropic virus (HTLV) types I and II by Western immunoblotting, polymerase chain reaction, and in situ hybridization of blood samples. The majority of patients were positive for HTLV antibodies by Western blotting and for HTLV-II gag sequences by polymerase chain reaction and in situ hybridization. Twenty nonexposure healthy controls were negative in all assays. These data support an association between an HTLV-II-like virus and CFIDS.

  17. Comparison of specimens for detection of porcine reproductive and respiratory syndrome virus infection in boar studs.

    PubMed

    Pepin, B J; Kittawornrat, A; Liu, F; Gauger, P C; Harmon, K; Abate, S; Main, R; Garton, C; Hargrove, J; Rademacher, C; Ramirez, A; Zimmerman, J

    2015-06-01

    Porcine reproductive and respiratory syndrome virus (PRRSV)-contaminated semen from boars is a route of transmission to females, and early detection of PRRSV infection in boars is a key component in sow farm biosecurity. The purpose of this study was to determine the optimum diagnostic specimen(s) for the detection of acute PRRSV infection in boars. Individually housed boars (n = 15) were trained for semen and oral fluid collection and then vaccinated with a commercial PRRSV modified live virus vaccine. Starting on the day of vaccination and for 14 days thereafter, oral fluid specimens were collected daily from all boars. The 15 boars were subdivided into three groups of 5, and serum, blood swabs and 'frothy saliva' were collected at the time of semen collection on a 3-day rotation. Frothy saliva, derived from the submandibular salivary gland, is produced by aroused boars. Semen was centrifuged, and semen supernatant and cell fractions were tested separately. All samples were randomly ordered and then tested by PRRSV real-time quantitative reverse-transcription polymerase chain reaction assay (rRT-PCR) and PRRSV antibody ELISA. In this study, a comparison of serum, blood swab, and oral fluid rRT-PCR results found no statistically significant differences in the onset of detection or proportion of positives, but serum was numerically superior to oral fluids for early detection. Serum and oral fluid provided identical rRT-PCR results at ? 5 day post-vaccination. Likewise, the onset of detection of PRRSV antibody in serum, oral fluid and frothy saliva was statistically equivalent, with serum results again showing a numerical advantage. These results showed that the highest assurance of providing PRRSV-negative semen to sow farms should be based on rRT-PCR testing of serum collected at the time of semen collection. This approach can be augmented with oral fluid sampling from a random selection of uncollected boars to provide for statistically valid surveillance of the boar stud. PMID:23895185

  18. Characterization of homologous and heterologous adaptive immune responses in porcine reproductive and respiratory syndrome virus infection

    PubMed Central

    2012-01-01

    The present study characterized the homologous and heterologous immune response in type-I porcine reproductive and respiratory syndrome virus (PRRSV) infection. Two experiments were conducted: in experiment 1, eight pigs were inoculated with PRRSV strain 3262 and 84?days post-inoculation (dpi) they were challenged with either strain 3262 or strain 3267 and followed for the next 14?days (98 dpi). In experiment 2, eight pigs were inoculated with strain 3267 and challenged at 84 dpi as above. Clinical course, viremia, humoral response (neutralizing and non-neutralizing antibodies, NA) and virus-specific IFN-? responses (ELISPOT) were evaluated all throughout the study. Serum levels of IL-1, IL-6, IL-8, TNF-? and TGF-? were determined (ELISA) after the second challenge. In experiment 1 primo-inoculation with strain 3262 induced viremia of???28?days, low titres of homologous NA but strong IFN-? responses. In contrast, strain 3267 induced longer viremias (up to 56?days), higher NA titres (? 6 log2) and lower IFN-? responses. Inoculation with 3267 produced higher serum IL-8 levels. After the re-challenge at 84 dpi, pigs in experiment 1 developed mostly a one week viremia regardless of the strain used. In experiment 2, neither the homologous nor the heterologous challenge resulted in detectable viremia although PRRSV was present in tonsils of some animals. Homologous re-inoculation with 3267 produced elevated TGF-? levels in serum for 7–14?days but this did not occur with the heterologous re-inoculation. In conclusion, inoculation with different PRRSV strains result in different virological and immunological outcomes and in different degrees of homologous and heterologous protection. PMID:22515169

  19. Inhibition of porcine reproductive and respiratory syndrome virus replication by RNA interference in MARC-145 cells.

    PubMed

    Bao, Yonghua; Guo, Yongchen; Zhang, Liying; Zhao, Zhihui; Li, Ning

    2012-03-01

    With the ultimate aim of producing an RNA interference-mediated transgenic pig that is resistant to porcine reproductive and respiratory syndrome virus (PRRSV), we have investigated the effect of RNA interference (RNAi) on silencing the expression of viral genes in the MARC-145 cell line. Twenty small interfering RNAs (siRNAs) were designed and screened for their ability to suppress the expression of the genes ORF1b, 5, 6, and 7 from the highly virulent isolate, PRRSV-JXwn06. Of these siRNAs, the four most effective were selected and four short hairpin RNA (shRNA) expression vectors (pGenesil-1-1b-135, pGenesil-1-1b-372, pGenesil-1-6-135, and pGenesil-1-6-169) targeting ORF1b and ORF6 were constructed and delivered into MARC-145 cells. These cells were then infected with JXwn06. All four vectors inhibited the PRRSV-specific cytopathic effect (CPE). The virus titers in cells transfected with pGenesil-1-1b-135, pGenesil-1-1b-372, pGenesil-1-6-135, and pGenesil-1-6-169 were lower than that of control cells by approximately 150-, 600-, 2.3- and 1.7-fold, respectively. In addition, the expression levels of ORF1 and ORF6 were reduced compared with controls. The unglycosylated membrane protein M, encoded by ORF6, was not detectable in cells transfected with shRNA expression vectors. These results verified that RNAi can effectively inhibit PRRSV-JXwn06 replication in cultured cells in vitro. The four shRNA expression vectors are an initial step in the production of transgenic pigs with PRRSV resistance. PMID:21667252

  20. Median infectious dose (ID??) of porcine reproductive and respiratory syndrome virus isolate MN-184 via aerosol exposure.

    PubMed

    Cutler, Timothy D; Wang, Chong; Hoff, Steven J; Kittawornrat, Apisit; Zimmerman, Jeffrey J

    2011-08-01

    The median infectious dose (ID(50)) of porcine reproductive and respiratory syndrome (PRRS) virus isolate MN-184 was determined for aerosol exposure. In 7 replicates, 3-week-old pigs (n=58) respired 10l of airborne PRRS virus from a dynamic aerosol toroid (DAT) maintained at -4°C. Thereafter, pigs were housed in isolation and monitored for evidence of infection. Infection occurred at virus concentrations too low to quantify by microinfectivity assays. Therefore, exposure dose was determined using two indirect methods ("calculated" and "theoretical"). "Calculated" virus dose was derived from the concentration of rhodamine B monitored over the exposure sequence. "Theoretical" virus dose was based on the continuous stirred-tank reactor model. The ID(50) estimate was modeled on the proportion of pigs that became infected using the probit and logit link functions for both "calculated" and "theoretical" exposure doses. Based on "calculated" doses, the probit and logit ID(50) estimates were 1 × 10(-0.13)TCID(50) and 1 × 10(-0.14)TCID(50), respectively. Based on "theoretical" doses, the probit and logit ID(50) were 1 × 10(0.26)TCID(50) and 1 × 10(0.24)TCID(50), respectively. For each point estimate, the 95% confidence interval included the other three point estimates. The results indicated that MN-184 was far more infectious than PRRS virus isolate VR-2332, the only other PRRS virus isolate for which ID(50) has been estimated for airborne exposure. Since aerosol ID(50) estimates are available for only these two isolates, it is uncertain whether one or both of these isolates represent the normal range of PRRS virus infectivity by this route. PMID:21474258

  1. Prevention and treatment of human immunodeficiency virus/acquired immunodeficiency syndrome in resource-limited settings.

    PubMed Central

    Hogan, Daniel R.; Salomon, Joshua A.

    2005-01-01

    Strategies for confronting the epidemic of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) have included a range of different approaches that focus on prevention and treatment. However, debate persists over what levels of emphasis are appropriate for the different components of the global response. This paper presents an overview of this debate and briefly summarizes the evidence on a range of interventions designed to prevent the spread of HIV infection, paying particular attention to voluntary counselling and testing, treatment for sexually transmitted infections and prevention of mother-to-child transmission. We also review the experience with antiretroviral therapy to date in terms of response rates and survival rates, adherence, drug resistance, behavioural change and epidemiological impact. Although various studies have identified strategies with proven effectiveness in reducing the risks of HIV infection and AIDS mortality, considerable uncertainties remain. Successful integration of treatment and prevention of HIV/AIDS will require a balanced approach and rigorous monitoring of the impact of programmes in terms of both individual and population outcomes. PMID:15744406

  2. Laguna Negra Virus Infection Causes Hantavirus Pulmonary Syndrome in Turkish Hamsters (Mesocricetus brandti).

    PubMed

    Hardcastle, K; Scott, D; Safronetz, D; Brining, D L; Ebihara, H; Feldmann, H; LaCasse, R A

    2016-01-01

    Laguna Negra virus (LNV) is a New World hantavirus associated with severe and often fatal cardiopulmonary disease in humans, known as hantavirus pulmonary syndrome (HPS). Five hamster species were evaluated for clinical and serologic responses following inoculation with 4 hantaviruses. Of the 5 hamster species, only Turkish hamsters infected with LNV demonstrated signs consistent with HPS and a fatality rate of 43%. Clinical manifestations in infected animals that succumbed to disease included severe and rapid onset of dyspnea, weight loss, leukopenia, and reduced thrombocyte numbers as compared to uninfected controls. Histopathologic examination revealed lung lesions that resemble the hallmarks of HPS in humans, including interstitial pneumonia and pulmonary edema, as well as generalized infection of endothelial cells and macrophages in major organ tissues. Histologic lesions corresponded to the presence of viral antigen in affected tissues. To date, there have been no small animal models available to study LNV infection and pathogenesis. The Turkish hamster model of LNV infection may be important in the study of LNV-induced HPS pathogenesis and development of disease treatment and prevention strategies. PMID:25722219

  3. [Development of human antibodies against the Gn protein of severe fever with thrombocytopenia syndrome virus].

    PubMed

    Chen, Suhua; Sun, Lina; Liu, Yang; Li, Chuan; Liu, Lin; Liang, Mifang; Qiu, Peihong

    2015-01-01

    To obtain human antibodies against the Gn protein of Severe fever with thrombocytopenia syndrome virus (SFTSV) with phage display technology, this study aimed to screen anti-Gn protein antibodies from an anti-SFTSV Fab human phage display library. Antibody genes were identified by sequence analysis and the specificity of antibodies was confirmed by ELISA. The Fab antibody genes were cloned into the HL51-14 vector and expressed in a mammalian cell expression system. IgG antibodies were then purified by protein A affinity chromatography,and the results were further confirmed by ELISA,IFA,western blotting assays and micro-neutralization tests. The results showed that, after three rounds of panning, there were 390 human Fab antibodies against SFTSV particles, of which 364 were specific for nucleoprotein. Coated with the Gn protein, eight different Fab antibodies specific for Gn protein were obtained after the determination of the subtype and subclass of antibodies by gene sequencing; five of these antibodies were from the Lambda library and three were from the Kappa library. The eight IgG antibodies could specifically bind to Gn protein according to the ELISA, IFA and Western blotting assays. The micro-neutralization test showed that these eight antibodies had no neutralizing activity,but they could still provide a reference for research in human monoclonal antibodies against SFTSV. PMID:25997326

  4. Probable aerosol transmission of severe fever with thrombocytopenia syndrome virus in southeastern China.

    PubMed

    Gong, Z; Gu, S; Zhang, Y; Sun, J; Wu, X; Ling, F; Shi, W; Zhang, P; Li, D; Mao, H; Zhang, L; Wen, D; Zhou, B; Zhang, H; Huang, Y; Zhang, R; Jiang, J; Lin, J; Xia, S; Chen, E; Chen, Z

    2015-12-01

    Some clusters of severe fever with thrombocytopenia syndrome virus (SFTSV) infection were reported in China as of 2010. However, to date, there has been no epidemiologic evidence of aerosol transmission of SFTSV. Epidemiologic investigations were conducted after a cluster of 13 cases of SFTSV in May 2014. A total of 13 cases, including 11 confirmed cases and one clinically diagnosed case, were identified besides the case of the index patient. The index patient experienced onset of SFTSV on 23 April and died on 1 May. The patients with secondary cases had onset from 10 to 16 May, peaking on 13 May. Moreover, eight secondary cases occurred in family members of the index patient, and the other five cases occurred in neighbors of the index patient. According to epidemiologic investigations, patients 1, 3, 4, 5, 6, 7, 9 and 12 contracted the disease through contact with blood of the index patient. Notably, patients 8 and 10 did not have a history of contact with the blood of the index patient, but they stayed in the mourning hall for hours. SFTSV could be transmitted from person to person by direct contact and/or aerosol transmission, and it is important to consider aerosol transmission as a possible transmission route. PMID:26255811

  5. Oral immunogenicity of porcine reproductive and respiratory syndrome virus antigen expressed in transgenic banana.

    PubMed

    Chan, Hui-Ting; Chia, Min-Yuan; Pang, Victor Fei; Jeng, Chian-Ren; Do, Yi-Yin; Huang, Pung-Ling

    2013-04-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a persistent threat of economically significant influence to the swine industry worldwide. Recombinant DNA technology coupled with tissue culture technology is a viable alternative for the inexpensive production of heterologous proteins in planta. Embryogenic cells of banana cv. 'Pei chiao' (AAA) have been transformed with the ORF5 gene of PRRSV envelope glycoprotein (GP5) using Agrobacterium-mediated transformation and have been confirmed. Recombinant GP5 protein levels in the transgenic banana leaves were detected and ranged from 0.021%-0.037% of total soluble protein. Pigs were immunized with recombinant GP5 protein by orally feeding transgenic banana leaves for three consecutive doses at a 2-week interval and challenged with PRRSV at 7 weeks postinitial immunization. A vaccination-dependent gradational increase in the elicitation of serum and saliva anti-PRRSV IgG and IgA was observed. Furthermore, significantly lower viraemia and tissue viral load were recorded when compared with the pigs fed with untransformed banana leaves. The results suggest that transgenic banana leaves expressing recombinant GP5 protein can be an effective strategy for oral delivery of recombinant subunit vaccines in pigs and can open new avenues for the production of vaccines against PRRSV. PMID:23116484

  6. Evolutionary diversification of type 2 porcine reproductive and respiratory syndrome virus.

    PubMed

    Brar, Manreetpal Singh; Shi, Mang; Murtaugh, Michael P; Leung, Frederick Chi-Ching

    2015-07-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the leading swine pathogens causing tremendous economic loss to the global swine industry due to its virulence, pathogenesis, infectivity and transmissibility. Although formally recognized only two and half decades ago, molecular dating estimation indicates a more ancient evolutionary history, which involved divergence into two genotypes (type 1 and type 2) prior to the 'initial' outbreaks of the late 1980s. Type 2 PRRSV circulates primarily in North America and Asia. The relatively greater availability of sequence data for this genotype from widespread geographical territories has enabled a better understanding of the evolving genotype. However, there are a number of challenges in terms of the vastness of data available and what this indicates in the context of viral diversity. Accordingly, here we revisit the mechanisms by which PRRSV generates variability, describe a means of organizing type 2 diversity captured in voluminous ORF5 sequences in a phylogenetic framework and provide a holistic view of known global type 2 diversity in the same setting. The consequences of the expanding diversity for control measures such as vaccination are discussed, as well as the contribution of modified live vaccines to the circulation of field isolates. We end by highlighting some limitations of current molecular epidemiology studies in relation to inferring PRRSV diversity, and what steps can be taken to overcome these and additionally enable PRRSV sequence data to be informative about viral phenotypic traits such as virulence. PMID:25711962

  7. [Terminology for classifying the porcine reproductive and respiratory syndrome virus (PRRSV) status of swine herds].

    PubMed

    Holtkamp, D J; Polson, D D; Torremorell, M; Morrison, B; Classen, D M; Becton, L; Henry, S; Rodibaugh, M T; Rowland, R R; Snelson, H; Straw, B; Yeske, P; Zimmerman, J

    2011-01-01

    Standardized terminology for the porcine reproductive and respiratory syndrome virus (PRRSV) status of swine herds is necessary to facilitate communication between veterinarians, swine producers, genetic companies, and other industry participants. It is also required for implementation of regional and national efforts towards PRRSV control and elimination. The purpose of this paper is to provide a herd classification system for describing the PRRSV status of herds, based upon a set of definitions reflecting the biology and ecology of PRRSV. The herd classification system was developed by a definitions committee formed jointly by the American Association of Swine Veterinarians (AASV) and the United States Department of Agriculture PRRS-Coordinated Agricultural Project, and was approved by the AASV Board of Directors on March 9, 2010. The committee included veterinarians from private practice and industry, researchers, and representatives from AASV and the National Pork Board. Breeding herds, with or without growing pigs on the same premises, are categorized as Positive Unstable (Category I), Positive Stable (Category II), Provisional Negative (Category III), or Negative (Category IV) on the basis of herd shedding and exposure status. Growing-pig herds are categorized as Positive or Negative. Recommended testing procedures and decision rules for herd classification are detailed. PMID:22138772

  8. Screening, isolation and optimization of anti–white spot syndrome virus drug derived from marine plants

    PubMed Central

    Chakraborty, Somnath; Ghosh, Upasana; Balasubramanian, Thangavel; Das, Punyabrata

    2014-01-01

    Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various marine floral ecosystems and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty species of marine plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti-WSSV property in Litopenaeus vannamei. By means of chemical processes, the purified anti-WSSV plant isolate, MP07X was derived. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug. Results Nine plant isolates exhibited significant survivability in host. The drug MP07X thus formulated showing 85% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of MP07X required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 1?000 mg/kg body weight/day survived at the rate of 85%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug MP07X derived from Rhizophora mucronata is a potent anti-WSSV drug. PMID:25183065

  9. Localization studies of two white spot syndrome virus structural proteins VP51 and VP76

    PubMed Central

    Wu, Chenglin; Yang, Feng

    2006-01-01

    VP51 and VP76 are two structural proteins of white spot syndrome virus (WSSV). However, there is some controversy about their localization in the virion at present. In this study, we employ multiple approaches to reevaluate the location of VP51 and VP76. Firstly, we found VP51 and VP76 presence in viral nucleocapsids fraction by Western blotting. Secondly, after the high-salt treatment of nucleocapsids, VP51 and VP76 were still exclusively present in viral capsids by Western blotting and immunoelectron microscopy, suggesting two proteins are structural components of the viral capsid. To gather more evidence, we developed a method based on immunofluorescence flow cytometry. The results revealed that the mean fluorescence intensity of the viral capsids group was significantly higher than that of intact virions group after incubation with anti-VP51 or anti-VP76 serum and fluorescein isothiocyanate conjugated secondary antibody. All these results indicate that VP51 and VP76 are both capsid proteins of WSSV. PMID:16968527

  10. Honeybee (Apis mellifera) Venom Reinforces Viral Clearance during the Early Stage of Infection with Porcine Reproductive and Respiratory Syndrome Virus through the Up-Regulation of Th1-Specific Immune Responses

    PubMed Central

    Lee, Jin-A; Kim, Yun-Mi; Hyun, Pung-Mi; Jeon, Jong-Woon; Park, Jin-Kyu; Suh, Guk-Hyun; Jung, Bock-Gie; Lee, Bong-Joo

    2015-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is a chronic and immunosuppressive viral disease that is responsible for substantial economic losses for the swine industry. Honeybee venom (HBV) is known to possess several beneficial biological properties, particularly, immunomodulatory effects. Therefore, this study aimed at evaluating the effects of HBV on the immune response and viral clearance during the early stage of infection with porcine reproductive and respiratory syndrome virus (PRRSV) in pigs. HBV was administered via three routes of nasal, neck, and rectal and then the pigs were inoculated with PRRSV intranasally. The CD4+/CD8+ cell ratio and levels of interferon (IFN)-? and interleukin (IL)-12 were significantly increased in the HBV-administered healthy pigs via nasal and rectal administration. In experimentally PRRSV-challenged pigs with virus, the viral genome load in the serum, lung, bronchial lymph nodes and tonsil was significantly decreased, as was the severity of interstitial pneumonia, in the nasal and rectal administration group. Furthermore, the levels of Th1 cytokines (IFN-? and IL-12) were significantly increased, along with up-regulation of pro-inflammatory cytokines (TNF-? and IL-1?) with HBV administration. Thus, HBV administration—especially via the nasal or rectal route—could be a suitable strategy for immune enhancement and prevention of PRRSV infection in pigs. PMID:26008237

  11. Generation of porcine reproductive and respiratory syndrome virus by in vitro assembly of viral genomic cDNA fragments.

    PubMed

    Suhardiman, Maman; Kramyu, Jarin; Narkpuk, Jaraspim; Jongkaewwattana, Anan; Wanasen, Nanchaya

    2015-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is the causative agent for a swine disease affecting the pig industry worldwide. Infection with PRRSV leads to reproductive complications, respiratory illness, and weak immunity to secondary infections. To better control PRRSV infection, novel approaches for generating control measures are critically needed. Here, in vitro Gibson assembly (GA) of viral genomic cDNA fragments was tested for its use as a quick and simple method to recover infectious PRRSV in cell culture. GA involves the activities of T5-exonuclease, Phusion polymerase, and Taq ligase to join overlapping cDNA fragments in an isothermal condition. Four overlapping cDNA fragments covering the entire PRRSV genome and one vector fragment were used to create a plasmid capable of expressing the PRRSV genome. The assembled product was used to transfect a co-culture of 293T and MARC-145 cells. Supernatants from the transfected cells were then passaged onto MARC-145 cells to rescue infectious virus particles. Verification and characterization of the recovered virus confirmed that the GA protocol generated infectious PRRSV that had similar characteristics to the parental virus. This approach was then tested for the generation of a chimeric virus. By replacing one of the four genomic fragments with that of another virus strain, a chimeric virus was successfully recovered via GA. In conclusion, this study describes for the first time the use of GA as a simple, yet powerful tool for generating infectious PRRSV needed for studying PRRSV biology and developing novel vaccines. PMID:25300804

  12. Successful vaccination strategies that protect aged mice from lethal challenge from influenza virus and heterologous severe acute respiratory syndrome coronavirus.

    PubMed

    Sheahan, Timothy; Whitmore, Alan; Long, Kristin; Ferris, Martin; Rockx, Barry; Funkhouser, William; Donaldson, Eric; Gralinski, Lisa; Collier, Martha; Heise, Mark; Davis, Nancy; Johnston, Robert; Baric, Ralph S

    2011-01-01

    Newly emerging viruses often circulate as a heterogeneous swarm in wild animal reservoirs prior to their emergence in humans, and their antigenic identities are often unknown until an outbreak situation. The newly emerging severe acute respiratory syndrome coronavirus (SARS-CoV) and reemerging influenza virus cause disproportionate disease in the aged, who are also notoriously difficult to successfully vaccinate, likely due to immunosenescence. To protect against future emerging strains, vaccine platforms should induce broad cross-reactive immunity that is sufficient to protect from homologous and heterologous challenge in all ages. From initial studies, we hypothesized that attenuated Venezuelan equine encephalitis virus (VEE) replicon particle (VRP) vaccine glycoproteins mediated vaccine failure in the aged. We then compared the efficacies of vaccines bearing attenuated (VRP(3014)) or wild-type VEE glycoproteins (VRP(3000)) in young and aged mice within novel models of severe SARS-CoV pathogenesis. Aged animals receiving VRP(3000)-based vaccines were protected from SARS-CoV disease, while animals receiving the VRP(3014)-based vaccines were not. The superior protection for the aged observed with VRP(3000)-based vaccines was confirmed in a lethal influenza virus challenge model. While the VRP(3000) vaccine's immune responses in the aged were sufficient to protect against lethal homologous and heterologous challenge, our data suggest that innate defects within the VRP(3014) platform mediate vaccine failure. Exploration into the mechanism(s) of successful vaccination in the immunosenescent should aid in the development of successful vaccine strategies for other viral diseases disproportionately affecting the elderly, like West Nile virus, influenza virus, norovirus, or other emerging viruses of the future. PMID:20980507

  13. Effect of the host cell line on the vaccine efficacy of an attenuated porcine reproductive and respiratory syndrome virus.

    PubMed

    Calzada-Nova, Gabriela; Husmann, Robert J; Schnitzlein, William M; Zuckermann, Federico A

    2012-07-15

    The abilities of the modified-live Prime Pac (PP) strain of porcine reproductive and respiratory syndrome virus (PRRSV), propagated in either traditional simian cells (MARC-145) or in a novel porcine alveolar macrophage cell line (ZMAC), to confer pigs protection against subsequent PRRSV challenge were compared. Eight week-old pigs were injected with PP virus grown in one of the two cell types and then exposed 4 weeks later to the "atypical" PRRSV isolate NADC-20. Control animals were similarly challenged or remained PRRSV-naïve. While the average adjusted body weight (aabw) of the strict control group increased 22% by 10 days post challenge (pc), this value for the non-vaccinated, challenged group dropped 4%. In contrast, prior immunization with PP virus, regardless of its host cell source, ameliorated this effect by affording a >9% rise in aabw. Likewise, nearly equivalent protection was extended to both groups of vaccinates in regards to the temporal elimination of their pc clinical distress and viremia. However, the PP virus propagated in ZMAC cells appeared to be more efficacious since four of the six pigs receiving this biologic cleared the challenge virus from the their lungs by 10 days pc as compared to only one member of the other vaccinated group. Notably, the predominant quasispecies in the ZMAC cell-prepared PP virus stock contained a highly conserved N-glycosylation site at position 184 in its glycoprotein 2 while this entity was underrepresented in the MARC-145 cell grown biologic. Since glycoprotein 2 is involved in infectivity, such additional glycosylation may enhance virus replication in porcine alveolar macrophages. PMID:22648044

  14. Detection of Porcine reproductive and respiratory syndrome virus infection in porcine oral fluid samples: a longitudinal study under experimental conditions.

    PubMed

    Prickett, John; Simer, Robert; Christopher-Hennings, Jane; Yoon, Kyoung-Jin; Evans, Richard B; Zimmerman, Jeffrey J

    2008-03-01

    Isolation of Porcine reproductive and respiratory syndrome virus (PRRSV) from oral fluids was first reported in 1997. The objective of the present study was to determine whether PRRSV and/or anti-PRRSV antibodies were present in oral fluids at diagnostic levels. The level and duration of PRRSV and anti-PRRSV antibodies in serum and oral fluids was evaluated in 3 age groups of pigs (4, 8, or 12 weeks of age) inoculated with a type 2 (North American) PRRSV isolate. Serum, buccal swabs, and pen-based oral fluid samples were collected for 63 days following inoculation. Specimens were assayed for PRRSV by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR), and for anti-PRRSV antibodies by enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT). Porcine reproductive and respiratory syndrome virus was detected by real-time qRT-PCR in serum for approximately 5 weeks and in oral fluids for approximately 4 weeks postinoculation. Pig age at the time of inoculation had no effect on the quantity or duration of virus in oral fluid samples. Low levels of anti-PRRSV antibody were detected in oral fluid samples by ELISA and IFAT. Although the approach remains to be validated in the field, the results of this experiment suggest that pen-based oral fluid sampling could be an efficient, cost-effective approach to PRRSV surveillance in swine populations. PMID:18319427

  15. Polarisation of Major Histocompatibility Complex II Host Genotype with Pathogenesis of European Brown Hare Syndrome Virus

    PubMed Central

    Iacovakis, Christos; Mamuris, Zissis; Moutou, Katerina A.; Touloudi, Antonia; Hammer, Anne Sofie; Valiakos, George; Giannoulis, Themis; Stamatis, Costas; Spyrou, Vassiliki; Athanasiou, Labrini V.; Kantere, Maria; Asferg, Tommy; Giannakopoulos, Alexios; Salomonsen, Charlotte M.; Bogdanos, Dimitrios; Birtsas, Periklis; Petrovska, Liljana; Hannant, Duncan; Billinis, Charalambos

    2013-01-01

    A study was conducted in order to determine the occurrence of European Brown Hare Syndrome virus (EBHSV) in Denmark and possible relation between disease pathogenesis and Major Histocompatibility Complex (MHC) host genotype. Liver samples were examined from 170 brown hares (hunted, found sick or dead), collected between 2004 and 2009. Macroscopical and histopathological findings consistent with EBHS were detected in 24 (14.1%) hares; 35 (20.6%) had liver lesions not typical of the syndrome, 50 (29.4%) had lesions in other tissues and 61 (35.9%) had no lesions. Sixty five (38.2%) of 170 samples were found to be EBHSV-positive (RT-PCR, VP60 gene). In order to investigate associations between viral pathogenesis and host genotype, variation within the exon 2 DQA gene of MHC was assessed. DQA exon 2 analysis revealed the occurrence of seven different alleles in Denmark. Consistent with other populations examined so far in Europe, observed heterozygosity of DQA (Ho?=?0.1180) was lower than expected (He?=?0.5835). The overall variation for both nucleotide and amino acid differences (2.9% and 14.9%, respectively) were lower in Denmark than those assessed in other European countries (8.3% and 16.9%, respectively). Within the peptide binding region codons the number of nonsynonymous substitutions (dN) was much higher than synonymous substitutions (dS), which would be expected for MHC alleles under balancing selection. Allele frequencies did not significantly differ between EBHSV-positive and -negative hares. However, allele Leeu-DQA*30 was detected in significantly higher (P?=?0.000006) frequency among the positive hares found dead with severe histopathological lesions than among those found sick or apparently healthy. In contrast, the latter group was characterized by a higher frequency of the allele Leeu-DQA*14 as well as the proportion of heterozygous individuals (P?=?0.000006 and P?=?0.027). These data reveal a polarisation between EBHSV pathogenesis and MHC class II genotype within the European brown hare in Denmark. PMID:24069299

  16. Expression, Purification, Crystallization of Two Major Envelope Proteins from White Spot Syndrome Virus

    SciTech Connect

    Tang,X.; Hew, C.

    2007-01-01

    White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapor-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 {angstrom} resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 {angstrom}. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 {angstrom}, and diffracts to 2.0 {angstrom} resolution.

  17. Reactomes of Porcine Alveolar Macrophages Infected with Porcine Reproductive and Respiratory Syndrome Virus

    PubMed Central

    Jiang, Zhihua; Zhou, Xiang; Michal, Jennifer J.; Wu, Xiao-Lin; Zhang, Lifan; Zhang, Ming; Ding, Bo; Liu, Bang; Manoranjan, Valipuram S.; Neill, John D.; Harhay, Gregory P.; Kehrli, Marcus E.; Miller, Laura C.

    2013-01-01

    Porcine reproductive and respiratory syndrome (PRRS) has devastated pig industries worldwide for many years. It is caused by a small RNA virus (PRRSV), which targets almost exclusively pig monocytes or macrophages. In the present study, five SAGE (serial analysis of gene expression) libraries derived from 0 hour mock-infected and 6, 12, 16 and 24 hours PRRSV-infected porcine alveolar macrophages (PAMs) produced a total 643,255 sequenced tags with 91,807 unique tags. Differentially expressed (DE) tags were then detected using the Bayesian framework followed by gene/mRNA assignment, arbitrary selection and manual annotation, which determined 699 DE genes for reactome analysis. The DAVID, KEGG and REACTOME databases assigned 573 of the DE genes into six biological systems, 60 functional categories and 504 pathways. The six systems are: cellular processes, genetic information processing, environmental information processing, metabolism, organismal systems and human diseases as defined by KEGG with modification. Self-organizing map (SOM) analysis further grouped these 699 DE genes into ten clusters, reflecting their expression trends along these five time points. Based on the number one functional category in each system, cell growth and death, transcription processes, signal transductions, energy metabolism, immune system and infectious diseases formed the major reactomes of PAMs responding to PRRSV infection. Our investigation also focused on dominant pathways that had at least 20 DE genes identified, multi-pathway genes that were involved in 10 or more pathways and exclusively-expressed genes that were included in one system. Overall, our present study reported a large set of DE genes, compiled a comprehensive coverage of pathways, and revealed system-based reactomes of PAMs infected with PRRSV. We believe that our reactome data provides new insight into molecular mechanisms involved in host genetic complexity of antiviral activities against PRRSV and lays a strong foundation for vaccine development to control PRRS incidence in pigs. PMID:23527143

  18. Porcine Reproductive and Respiratory Syndrome Virus Induces Interleukin-15 through the NF-?B Signaling Pathway

    PubMed Central

    Fu, Yi; Quan, Rong; Zhang, Hexiao; Hou, Jun

    2012-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) mainly infects macrophages/dendritic cells and modulates cytokine expression in these cells. Interleukin-15 (IL-15) is a pleiotropic cytokine involved in wide range of biological activities. It has been shown to be essential for the generation, activation, and proliferation of NK and NKT cells and for the survival and activation of CD8+ effector and memory T cells. In this study, we discovered that PRRSV infection upregulated IL-15 production at both the mRNA and protein levels in porcine alveolar macrophages (PAMs), blood monocyte-derived macrophages (BMo), and monocyte-derived dendritic cells (DCs). We subsequently demonstrated that the NF-?B signaling pathway was essential for PRRSV infection-induced IL-15 production. First, addition of an NF-?B inhibitor drastically reduced PRRSV infection-induced IL-15 production. We then found that NF-?B was indeed activated upon PRRSV infection, as evidenced by I?B phosphorylation and degradation. Moreover, we revealed an NF-?B binding motif in the cloned porcine IL-15 (pIL-15) promoter, deletion of which abrogated the pIL-15 promoter activity in PRRSV-infected alveolar macrophages. In addition, we demonstrated that PRRSV nucleocapsid (N) protein had the ability to induce IL-15 production in porcine alveolar macrophage cell line CRL2843 by transient transfection, which was mediated by its multiple motifs, and it also activated NF-?B. These data indicated that PRRSV infection-induced IL-15 production was likely through PRRSV N protein-mediated NF-?B activation. Our findings provide new insights into the molecular mechanisms underling the IL-15 production induced by PRRSV infection. PMID:22573868

  19. Efficacy of Fostera PRRS modified live virus vaccine against a Canadian heterologous virulent field strain of porcine reproductive and respiratory syndrome virus.

    PubMed

    Savard, Christian; Alvarez, Fernando; Provost, Chantale; Chorfi, Younes; D'Allaire, Sylvie; Benoit-Biancamano, Marie-Odile; Gagnon, Carl A

    2016-01-01

    Vaccination is a useful option to control infection with porcine reproductive and respiratory syndrome virus (PRRSV), and several modified live-PRRSV vaccines have been developed. These vaccines have shown some efficacy in reducing the incidence and severity of clinical disease as well as the duration of viremia and virus shedding but have failed to provide sterilizing immunity. The efficacy of modified live-virus (MLV) vaccines is greater against a homologous strain compared with heterologous PRRSV strains. The objective of this study was to evaluate the efficacy of Fostera PRRS MLV vaccine in protecting against challenge with a heterologous field strain widely circulating in the swine herds of eastern Canada. Forty-six piglets were divided into 4 groups: nonvaccinated-nonchallenged; nonvaccinated-challenged; vaccinated-challenged; and vaccinated-nonchallenged. The animals were vaccinated at 23 d of age with Fostera PRRS and challenged 23 d later with a heterologous field strain of PRRSV (FMV12-1425619). Overall, the vaccine showed some beneficial effects in the challenged animals by reducing the severity of clinical signs and the viral load. A significant difference between nonvaccinated and vaccinated animals was detected for some parameters starting 11 to 13 d after challenge, which suggested that the cell-mediated immune response or other delayed responses could be more important than pre-existing PRRSV antibodies in vaccinated animals within the context of protection against heterologous strains. PMID:26732457

  20. Porcine reproductive and respiratory syndrome virus: antigenic and molecular diversity of British isolates and implications for diagnosis.

    PubMed

    Frossard, Jean-Pierre; Fearnley, Catherine; Naidu, Brindha; Errington, Jane; Westcott, David G; Drew, Trevor W

    2012-08-17

    Porcine reproductive and respiratory syndrome (PRRS) is an endemic disease of pigs, caused by PRRS virus, a member of the Arteriviridae family. First seen in Britain in 1991, the disease continues to be a significant economic and welfare problem for pig producers. To date, only PRRSV genotype 1 has been found in Britain. At the genetic level, a considerable increase has been reported in the diversity of PRRS viruses isolated in Britain between 2003 and 2007, versus the early 1990 s. In this study, the diversity has been shown to extend to the antigenic level too, with potential consequences for diagnostic methods. Antigenic diversity was assessed using a panel of twelve monoclonal antibodies, only one of which reacted with all isolates tested. Nine diverse viruses were compared as potential antigens in immunoperoxidase monolayer assays, where each one produced quite different results for a common panel of sera. As a single virus is used in each diagnostic assay, results must therefore be interpreted cautiously. For a real-time RT-PCR assay, published oligonucleotide primer and probe sequences were evaluated against available genetic sequences of British and European viruses, and were re-designed where considerable mismatches were found. The multiplex assay incorporating these modified primers to detect genotype 1 and 2 PRRS viruses was then validated for use with diagnostic sera and tissues. As the increasing degree of diversity exhibited by British strains is mirrored in other countries, PRRSV will continue to provide an ongoing challenge to diagnosis at a global, as well as national level. PMID:22472704

  1. Detection of a pneumonia virus of mice (PVM) in an African hedgehog (Atelerix arbiventris) with suspected wobbly hedgehog syndrome (WHS).

    PubMed

    Madarame, Hiroo; Ogihara, Kikumi; Kimura, Moe; Nagai, Makoto; Omatsu, Tsutomu; Ochiai, Hideharu; Mizutani, Tetsyuya

    2014-09-17

    A pneumonia virus of mice (PVM) from an African hedgehog (Atelerix arbiventris) with suspected wobbly hedgehog syndrome (WHS) was detected and genetically characterized. The affected hedgehog had a nonsuppurative encephalitis with vacuolization of the white matter, and the brain samples yielded RNA reads highly homogeneous to PVM strain 15 (96.5% of full genomic sequence homology by analysis of next generation sequencing). PVM antigen was also detected in the brain and the lungs immunohistochemically. A PVM was strongly suggested as a causative agent of encephalitis of a hedgehog with suspected WHS. This is a first report of PVM infection in hedgehogs. PMID:25129384

  2. Epstein-Barr and human immunodeficiency viruses in acquired immunodeficiency syndrome-related primary central nervous system lymphoma.

    PubMed Central

    Morgello, S.

    1992-01-01

    The prevalence of Epstein-Barr virus (EBV) and human immunodeficiency virus (HIV) in acquired immunodeficiency syndrome (AIDS)-related primary central nervous system (CNS) lymphoma was examined. Deoxyribonucleic acid (DNA) extracted from 12 formalin-fixed, paraffin-embedded tumors was used as substrate for the polymerase chain reaction (PCR). Targets for amplification were the EBNA-1 region of EBV, the gag region of HIV, and a single copy cellular sequence as a control. The cases studied were autopsy and surgical specimens collected between the years 1985 and 1989. By the working formulation for non-Hodgkin's lymphomas, five had large cell, four had mixed large and small cleaved cell, two had small cleaved cell, and one had an unclassified histology. Epstein-Barr virus was detected in 6 of 12 tumors studied. Human immunodeficiency virus was not detected in any of the tumors. The presence of EBV was not correlated with any particular histologic tumor type. It is concluded that EBV, not HIV, can be detected in a large percentage (50%) of AIDS-related primary central nervous system (CNS) lymphomas. This viral association may be significant in light of the demonstrated ability of EBV to induce lymphoid tumors in experimental mammalian systems. Images Figure 1 Figure 2 PMID:1323221

  3. An aqueous platinum nanotube based fluorescent immuno-assay for porcine reproductive and respiratory syndrome virus detection.

    PubMed

    Chen, Lu; Ye, Shiyi; Cai, Kai; Zhang, Cuiling; Zhou, Guohua; He, Zhike; Han, Heyou

    2015-11-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) has been a significant pathogen towards global swine industry upon its emergence in the late 1980s and since then has exemplified a rapidly evolving, widely spreading pathogen. It is urgently important to develop a simple, rapid and cost effective method to detect this pathogen when virus outbreaks. In the present work, it was found that virus antibody modified platinum nanotubes (Pt-Ab) could act as a superquencher to CdTe:Zn(2+) quantum dots (CdTe:Zn(2+) QDs) fluorescence by Stern-Volmer constants nearly 10(9)M(-1) without any aggregation, the CdTe:Zn(2+) QDs fluorescence will recover as the Pt-Ab goes away by antibody and antigen interaction when virus was added into the probe solution, releasing CdTe:Zn(2+) QDs from the surface of Pt-Ab. By the recovery fluorescence intensity, it can realize qualitative and quantitative detection of PRRSV. This method gives a fast response to PRRSV concentration and provides a sensitive detection limit (2.4ng/mL). Moreover, it can be applied in infected porcine serum samples and obtain satisfied results. PMID:26452829

  4. Experimental and Natural Infections of Goats with Severe Fever with Thrombocytopenia Syndrome Virus: Evidence for Ticks as Viral Vector

    PubMed Central

    Han, Yewu; Guo, Xiling; Shi, Zhiyang; Wang, Hua; Zhou, Minghao

    2015-01-01

    Background Severe fever with thrombocytopenia syndrome virus (SFTSV), the causative agent for the fatal life-threatening infectious disease, severe fever with thrombocytopenia syndrome (SFTS), was first identified in the central and eastern regions of China. Although the viral RNA was detected in free-living and parasitic ticks, the vector for SFTSV remains unsettled. Methodology/Principal Findings Firstly, an experimental infection study in goats was conducted in a bio-safety level-2 (BSL-2) facility to investigate virus transmission between animals. The results showed that infected animals did not shed virus to the outside through respiratory or digestive tract route, and the control animals did not get infected. Then, a natural infection study was carried out in the SFTSV endemic region. A cohort of naïve goats was used as sentinel animals in the study site. A variety of daily samples including goat sera, ticks and mosquitoes were collected for viral RNA and antibody (from serum only) detection, and virus isolation. We detected viral RNA from free-living and parasitic ticks rather than mosquitoes, and from goats after ticks’ infestation. We also observed sero-conversion in all members of the animal cohort subsequently. The S segment sequences of the two recovered viral isolates from one infected goat and its parasitic ticks showed a 100% homology at the nucleic acid level. Conclusions/Significance In our natural infection study, close contact between goats does not appear to transmit SFTSV, however, the naïve animals were infected after ticks’ infestation and two viral isolates derived from an infected goat and its parasitic ticks shared 100% of sequence identity. These data demonstrate that the etiologic agent for goat cohort’s natural infection comes from environmental factors. Of these, ticks, especially the predominant species Haemaphysalis longicornis, probably act as vector for this pathogen. The findings in this study may help local health authorities formulate and focus preventive measures to contain this infection. PMID:26485390

  5. A Single Amino Acid Deletion in the Matrix Protein of Porcine Reproductive and Respiratory Syndrome Virus Confers Resistance to a Polyclonal Swine Antibody with Broadly Neutralizing Activity

    PubMed Central

    Popescu, Luca N.; Monday, Nicholas; Calvert, Jay G.; Rowland, Raymond R. R.

    2015-01-01

    Assessment of virus neutralization (VN) activity in 176 pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV) identified one pig with broadly neutralizing activity. A Tyr-10 deletion in the matrix protein provided escape from broad neutralization without affecting homologous neutralizing activity. The role of the Tyr-10 deletion was confirmed through an infectious clone with a Tyr-10 deletion. The results demonstrate differences in the properties and specificities of VN responses elicited during PRRSV infection. PMID:25855739

  6. Epidemiology of acquired immunodeficiency syndrome and human immunodeficiency virus infection in adolescents.

    PubMed

    Gayle, H D; D'Angelo, L J

    1991-04-01

    The epidemiology of acquired immunodeficiency syndrome (AIDS) and human immunodeficiency virus infection (HIV) in adolescents is important for purposes of prevention and car, since sexual and drug behavior is formed during this period. For 1990 the Centers for Disease Control reports .4% of AIDS cases are among adolescents 13-19 years old; this figure has steadily risen since 1982. 53% of the reported AIDS cases were from New York, Florida, California, Texas, Puerto Rico, and New Jersey, and has remained stable since 1984. 72% were from metropolitan areas of 1 million population, with a small decreasing trend between 1986-88. 75% of reported cases occurred between 17-19 years, and usually among males (80%) and ethnic minorities (36% African Americans and 18% Hispanics). The sex ratio dropped from 4:1 to 3:1 in 1988. Modes of transmission; indicator diseases and mortality; HIV seroprevalence data; risk of HIV transmission in adolescents; knowledge, attitudes, beliefs, and behavior; research needs; and prevention are discussed. From the studies available, it is reported that adolescents are aware that sexual intercourse and sharing IV drug needles are the main modes of HIV transmission. HIV transmission is more likely to be associated with homosexual contact. Misconceptions are that one could tell if a person were infected with HIV. Knowledge does not always translate to appropriate behavior. Perceived risk does decrease risky behavior, i.e., through abstinence or condom use. More information was desired. Research needs were identified as lagging behind present knowledge of children and adults, and necessary in clinical, epidemiologic, behavioral, and prevention aspects. The natural history of infection is limited to studies of hemophilia, where infected adolescents may have a lower rate of progression to AIDS or a longer incubation period or higher tolerance to severe immunodeficiency. Questions arise concerning the unique factors, such as hormonal changes, that influence the clinical course of the infection. Health care models need to be assessed. Identification of subpopulations that are at the highest risk is needed, i.e., the influence of the crack cocaine epidemic on HIV transmission. Prevention is seen in terms of new creative approaches, comprehensive school and nonschool health education, and behavioral techniques to avoid risky behavior throughout the health community. PMID:2062630

  7. Does varicella-zoster virus infection of the peripheral ganglia cause Chronic Fatigue Syndrome?

    PubMed

    Shapiro, Judith S

    2009-11-01

    This article posits that infection of the peripheral ganglia causes at least some cases of Chronic Fatigue Syndrome (CFS), with a neurotropic herpesvirus, particularly varicella-zoster virus (VZV), as the most likely cause of the infection. Virtually all CFS symptoms could be produced by an infection of the peripheral ganglia, with infection of the autonomic ganglia causing fatigue, postural hypotension, and sleep disturbances, and infection of the sensory ganglia causing sensory symptoms such as chronic pain. Furthermore, infections of the peripheral ganglia are known to cause long-term nerve dysfunction, which would help explain the chronic course of CFS. Herpesviruses have long been suspected as the cause of CFS; this theory has recently been supported by studies showing that administering antiherpes agents causes substantial improvement in some CFS patients. VZV is known to frequently reactivate in the peripheral ganglia of previously healthy adults and cause sudden, debilitating illness, making it a likely candidate as a cause of CFS. Moreover, many of the symptoms of CFS overlap with those of herpes zoster (shingles), with the exception that painful rash is not one of the symptoms of CFS. A model is therefore proposed in which CFS is one of the many manifestations of zoster sine herpete; that is, herpes zoster without rash. Furthermore, re-exposure to VZV in the form of chickenpox has become less common in the past few decades; without such re-exposure, immunity to VZV drops, which could explain the increased incidence of CFS. Co-infection with multiple herpesviruses is a possibility, as some CFS patients show signs of infection with other herpesviruses including Epstein-Barr, Cytomegalovirus, and HHV6. These three herpesviruses can attack immune cells, and may therefore promote neurotropic herpesvirus reactivation in the ganglia. The possibility of VZV as the causal agent in CFS has previously received almost no attention; the possibility that CFS involves infection of the peripheral ganglia has likewise been largely overlooked. This suggests that the search for a viral cause of CFS has been far from exhaustive. Several antiherpes drugs are available, as is a vaccine for VZV; more research into such agents as possible treatments for CFS is urgently needed. PMID:19520522

  8. Effect of porcine reproductive and respiratory syndrome virus on porcine alveolar macrophage function as determined using serial analysis of gene expression (SAGE)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine worldwide and causes considerable economic loss. The main target of infection is the porcine alveolar macrophage (PAM). Infection of PAM by PRRSV causes significant changes in their function by mechanisms that a...

  9. In depth global analysis of transcript abundance levels in porcine alveolar macrophages following infection with porcine reproductive and respiratory syndrome virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine worldwide and causes considerable economic loss. Infection of the primary target cells, porcine alveolar macrophages (PAMs), by PRRSV causes significant changes in their function by mechanisms that are not under...

  10. Changes in leukocyte subsets of pregnant gilts experimentally infected with porcine reproductive and respiratory syndrome virus and relationships with viral load and fetal outcome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In spite of more than two decades of extensive research, the understanding of porcine reproductive and respiratory syndrome virus (PRRSv) immunity is still incomplete. A PRRSv infection of the late term pregnant female can result in abortions, early farrowings, fetal death, and the birth of weak, co...

  11. Complete Genome Sequence of a Novel Natural Recombinant Porcine Reproductive and Respiratory Syndrome Virus Isolated from a Pig Farm in Yunnan Province, Southwest China

    PubMed Central

    Yan, Yulin; Xin, Aiguo; Zhu, Gaohong; Huang, Hui; Liu, Qian; Shao, Zhiyong; Zang, Yating; Chen, Ling; Sun, Yongke

    2013-01-01

    YN-2011 is a highly pathogenic North American porcine reproductive and respiratory syndrome virus (PRRSV). Unlike previously described PRRSVs, which contained a 30-amino-acid deletion in NS2, YN-2011 had no amino acid deletions or insertions but had several new mutations in NS2. Here, we announce the complete genome sequence of YN-2011. PMID:23405309

  12. Swine tracheobronchial lymph node mRNA responses in swine infected with a highly pathogenic strain of porcine reproductive and respiratory syndrome virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine worldwide. Emergence in 2006 of a novel highly pathogenic PRRSV (HP-PRRSV) isolate in China necessitated a comparative investigation into the host transcriptome response in tracheobronchial lymph nodes (TBLN) 14...

  13. Effect of porcine reproductive and respiratory syndrome virus on porcine alveolar macrophage function as determined using serial analysis of gene expression (SAGE)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major pathogen of swine worldwide and causes considerable economic loss. The major target of infection is the alveolar macrophage (AM). Infection of AMs by PRRSV causes significant changes in their function by mechanisms that are not...

  14. Validation of a major quantitative trait locus associated with host response to experimental infection with Porcine Reproductive and Respiratory Syndrome virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infectious diseases are costly to the swine industry and porcine reproductive and respiratory syndrome virus (PRRSV) is the most devastating. In earlier work, a quantitative trait locus associated with resistance/susceptibility to PRRSV was identified on Sus scrofa chromosome 4 (SSC4) using ~560 exp...

  15. A Critical Review of Human Immunodeficiency Virus Infection--And Acquired Immunodeficiency Syndrome-Related Research: The Knowledge, Attitudes, and Practice of Nurses.

    ERIC Educational Resources Information Center

    Swanson, Janice M.; And Others

    1990-01-01

    Reviews the research literature related to nurses' knowledge, attitudes, and practices concerning acquired immunodeficiency syndrome (AIDS), human immunodeficiency virus infection, and care of people with AIDS. Gaps in knowledge and negative, fearful attitudes were identified; negative fears and attitudes decreased with the gain in accurate…

  16. Porcine Reproductive and Respiratory Syndrome Virus Replicase - Isoforms of Nonstructural Protein 2 and Interaction with Heat Shock 70kDa Protein 5

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The nsp2 replicase protein of porcine reproductive and respiratory syndrome virus (PRRSV), when expressed independently, was recently demonstrated to be processed from its precursor by the PL2 protease at or near the G**1196|G**1197 dipeptide in transfected CHO cells. The proteolytic cleavage of nsp...

  17. Inhibition of highly pathogenic porcine reproductive and respiratory syndrome virus replication by recombinant pseudorabies virus-mediated RNA interference in piglets.

    PubMed

    Cao, Su-Fang; Guo, Qing-Yong; Wang, Yan

    2015-12-31

    Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is a variant of porcine reproductive and respiratory syndrome virus (PRRSV) which, in recent years, has caused heavy economic losses to swine-producing areas. Although current vaccines are somewhat prophylactic, they provide only limited protection. Furthermore, there are currently no effective anti-HP-PRRSV drugs. Consequently, it is necessary to develop novel antiviral strategies. In the present study, three recombinant pseudorabies viruses (PRV) expressing siRNAs against the ORF7 of HP-PRRSV strain HN1 (PRV gG-/siRNAN1, PRV gG-/siRNAN2, and PRV gG-/siRNAN3) were evaluated for the inhibition of HP-PRRSV replication. The results indicated that recombinant PRV-mediated siRNA could significantly decrease the replication of traditional PRRSV strain H1 at mRNA and protein levels in Marc-145 cells. Moreover, one recombinant PRV (PRV gG-/siRNAN2) was found to be inhibit the multiplication of HP-PRRSV strain HN1 effectively in Marc-145 cells at both the protein and ORF7 mRNA level. Twenty 21-day-old healthy weaned piglets were divided into four groups of five piglets each. Groups 1 and 2 were injected i.m. with PRV gG-/siRNAN2 and PRV gG-/siRNANeg individually. The piglets in group 3 were challenged with the HP-PRRSV control. After 24h, the piglets in groups 1-3 were challenged i.m. with HP-PRRSV strain HN1, while those in group 4 were i.m. administered with PBS as a negative control. The results showed that HP-PRRSV in serum and lung samples from piglets was effectively inhibited by PRV gG-/siRNAN2. The clinical signs and gross lesions of piglets inoculated with PRV gG-/siRNAN2 were significantly less invasive than those of the PRV gG-/siRNANeg group and HP-PRRSV control group. These results showed that siRNAs mediated by recombinant PRV could effectively suppress HP-PRRSV replication in vitro as well as in vivo. RNAi mediated by recombinant PRV presents a potential novel method to prevent HP-PRRSV infections in swine. However, the protective efficiency of PRV gG-/siRNAN2 should be assessed in a larger number of piglets in future studies. PMID:26560709

  18. Immune response and persistence of the porcine reproductive and respiratory syndrome virus in infected pigs and farm units.

    PubMed

    Albina, E; Madec, F; Cariolet, R; Torrison, J

    1994-05-28

    The kinetics of the serum antibody response to porcine reproductive and respiratory syndrome (PRRS) virus and the persistence of the virus after infection were determined in experimentally and naturally infected pigs. In an experimental study, four specific pathogen free (SPF) sows were infected with a French strain of PRRS virus at 90 days of gestation, and their piglets (the test piglets) were monitored for 29 weeks from birth. In one litter, antibodies against PRRS virus were absent before the piglets had ingested colostrum. Four days after birth, passive antibodies were present in the serum of these piglets, but they had disappeared by three weeks (just before weaning) when clinical signs were observed in a minority of the pigs. In a second litter, most of the piglets had no detectable antibodies until they were four weeks old, and clinical signs were observed during their second week of life. By eight weeks, antibodies were detected in all the pigs, and they persisted until observations ceased at 29 weeks. Two groups of three SPF pigs were placed in direct contact with the test piglets when they were four weeks old and a group of five SPF pigs was placed in indirect contact when they were 13 weeks old. The first two groups showed clinical signs and seroconverted but the third group did not. At 22 weeks old, two of the test piglets were subjected to movement stress and were given exogenous corticosteroids, after which the in-contact SPF pigs developed clinical signs and seroconverted.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7941249

  19. Schizophrenia or possession?

    PubMed

    Irmak, M Kemal

    2014-06-01

    Schizophrenia is typically a life-long condition characterized by acute symptom exacerbations and widely varying degrees of functional disability. Some of its symptoms, such as delusions and hallucinations, produce great subjective psychological pain. The most common delusion types are as follows: "My feelings and movements are controlled by others in a certain way" and "They put thoughts in my head that are not mine." Hallucinatory experiences are generally voices talking to the patient or among themselves. Hallucinations are a cardinal positive symptom of schizophrenia which deserves careful study in the hope it will give information about the pathophysiology of the disorder. We thought that many so-called hallucinations in schizophrenia are really illusions related to a real environmental stimulus. One approach to this hallucination problem is to consider the possibility of a demonic world. Demons are unseen creatures that are believed to exist in all major religions and have the power to possess humans and control their body. Demonic possession can manifest with a range of bizarre behaviors which could be interpreted as a number of different psychotic disorders with delusions and hallucinations. The hallucination in schizophrenia may therefore be an illusion-a false interpretation of a real sensory image formed by demons. A local faith healer in our region helps the patients with schizophrenia. His method of treatment seems to be successful because his patients become symptom free after 3 months. Therefore, it would be useful for medical professions to work together with faith healers to define better treatment pathways for schizophrenia. PMID:23269538

  20. A Novel Detection Platform for Shrimp White Spot Syndrome Virus Using an ICP11-Dependent Immunomagnetic Reduction (IMR) Assay

    PubMed Central

    Liu, Bing-Hsien; Lin, Yu-Chen; Ho, Chia-Shin; Yang, Che-Chuan; Chang, Yun-Tsui; Chang, Jui-Feng; Li, Chun-Yuan; Cheng, Cheng-Shun; Huang, Jiun-Yan; Lee, Yen-Fu; Hsu, Ming-Hung; Lin, Feng-Chun; Wang, Hao-Ching; Lo, Chu-Fang; Yang, Shieh-Yueh; Wang, Han-Ching

    2015-01-01

    Shrimp white spot disease (WSD), which is caused by white spot syndrome virus (WSSV), is one of the world’s most serious shrimp diseases. Our objective in this study was to use an immunomagnetic reduction (IMR) assay to develop a highly sensitive, automatic WSSV detection platform targeted against ICP11 (the most highly expressed WSSV protein). After characterizing the magnetic reagents (Fe3O4 magnetic nanoparticles coated with anti ICP11), the detection limit for ICP11 protein using IMR was approximately 2 x 10?3 ng/ml, and the linear dynamic range of the assay was 0.1~1 x 106 ng/ml. In assays of ICP11 protein in pleopod protein lysates from healthy and WSSV-infected shrimp, IMR signals were successfully detected from shrimp with low WSSV genome copy numbers. We concluded that this IMR assay targeting ICP11 has potential for detecting the WSSV. PMID:26380977

  1. Emergence of a virulent porcine reproductive and respiratory syndrome virus in vaccinated herds in the United States.

    PubMed

    Wang, Xiong; Marthaler, Douglas; Rovira, Albert; Rossow, Stephanie; Murtaugh, Michael P

    2015-12-01

    In early 2014, a Minnesota sow farm with a solid vaccination history suffered a severe porcine reproductive and respiratory syndrome (PRRS) outbreak with unusually high morbidity and mortality in piglets and sows, as well as anorexia and secondary bacterial infections in nursery pigs. Due to the unusual clinical severity in a PRRS-immune herd, genetic characteristics of the virus were examined to determine if a new PRRSV genotype had emerged. Phylogenetic analysis indicated that the virulent strain (PRRSV2/USA/Minnesota414/2014) was related to virulent strains circulating in the mid-western United States in recent years, and that the nonstructural protein 2 (nsp2) gene of MN414 contained an insertion-deletion pattern typical of existing type 2 virulent strains. We conclude that the MN414 isolate is a recently evolved member of the virulent lineage 1 family of type 2 PRRSV. PMID:26169029

  2. Generation and immunogenicity of transgenic potato expressing the GP5 protein of porcine reproductive and respiratory syndrome virus.

    PubMed

    Chen, Xia; Liu, Jinlin

    2011-04-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is an important pathogen that has caused huge economic losses in the global swine industry. The advent of molecular farming has provided a cost-effective strategy for the development of transgenic plants as bioreactors to produce recombinant proteins. In this study, transgenic potato expressing GP5 protein of PRRSV was produced by Agrobacterium-mediated transformation, and confirmed using Southern blot and RT-PCR analyses. Recombinant GP5 protein was detected by ELISA and Western blot analyses. Mice immunized with transgenic potato extracts generated both serum and gut mucosal-specific antibodies, although low levels of neutralizing antibodies were elicited. This study provides a new approach for the production of vaccines against PRRSV. PMID:21300109

  3. Identification and characterization of a prawn white spot syndrome virus gene that encodes an envelope protein VP31

    SciTech Connect

    Li Li; Xie Xixian; Yang Feng . E-mail: mbiotech@public.xm.fj.cn

    2005-09-15

    Based on a combination of SDS-PAGE and mass spectrometry, a protein with an apparent molecular mass of 31 kDa (termed as VP31) was identified from purified shrimp white spot syndrome virus (WSSV) envelope fraction. The resulting amino acid (aa) sequence matched an open reading frame (WSV340) of the WSSV genome. This ORF contained 783 nucleotides (nt), encoding 261 aa. A fragment of WSV340 was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein with a 6His-tag, and then specific antibody was raised. Western blot analysis and the immunoelectron microscope method (IEM) confirmed that VP31 was present exclusively in the viral envelope fraction. The neutralization experiment suggested that VP31 might play an important role in WSSV infectivity.

  4. Novel porcine reproductive and respiratory syndrome virus strains in the United States with deletions in untranslated regions.

    PubMed

    Wang, Leyi; Zhang, Yan

    2015-12-01

    Porcine reproductive and respiratory syndrome (PRRS) still causes major problems for the swine industry worldwide. Here, we report the detection and genomic characterization of two novel PRRS virus (PRRSV) strains from the United States with deletions in untranslated regions (UTRs). The OH155-2015 strain has two single-nucleotide deletions in the 5' UTR, whereas the OH28372-2013 strain has a 13-nt deletion in the 3' UTR. In addition, OH155-2015 and OH28372-2013 have a unique deletion and mutations in the NSP2 and N gene, respectively. Our study highlights the importance of continued monitoring of PRRSV using whole-genome sequencing. PMID:26358265

  5. Immunohistochemical characterization of type II pneumocyte proliferation after challenge with type I porcine reproductive and respiratory syndrome virus.

    PubMed

    Balka, G; Ladinig, A; Ritzmann, M; Saalmüller, A; Gerner, W; Käser, T; Jakab, C; Rusvai, M; Weißenböck, H

    2013-01-01

    The aim of this study was to characterize histologically and immunohistochemically the lung lesions developing in growing pigs, 10 and 21 days after experimental challenge with a field strain of porcine reproductive and respiratory syndrome virus (PRRSV). Lung lesions were scored for (1) pneumocyte hypertrophy and hyperplasia, (2) septal mononuclear infiltration, (3) intra-alveolar necrotic debris, (4) intra-alveolar inflammatory cell accumulation and (5) perivascular inflammatory cell accumulation. Immunohistochemistry was performed using antibodies specific for cytokeratin, Ki67, thyroid transcription factor (TTF)-1, the myelomonocytic marker MAC387 and PRRSV. Anti-TTF-1 identified type II pneumocytes and there was marked proliferation of these cells compared with control lung (P <0.05). Anti-cytokeratin labelled type I and II pneumocytes as well as bronchial epithelial cells; however, this labelling was not suitable for cell counting purposes. There was a correlation between lesion severity and the number of cells expressing Ki67 (P <0.05). PMID:23453491

  6. A model for the dynamic nuclear/nucleolar/cytoplasmic trafficking of the porcine reproductive and respiratory syndrome virus (PRRSV) nucleocapsid protein based on live cell imaging

    SciTech Connect

    You, Jae-Hwan; Howell, Gareth; Pattnaik, Asit K.; Osorio, Fernando A.; Hiscox, Julian A.

    2008-08-15

    Porcine reproductive and respiratory syndrome virus (PRRSV), an arterivirus, in common with many other positive strand RNA viruses, encodes a nucleocapsid (N) protein which can localise not only to the cytoplasm but also to the nucleolus in virus-infected cells and cells over-expressing N protein. The dynamic trafficking of positive strand RNA virus nucleocapsid proteins and PRRSV N protein in particular between the cytoplasm and nucleolus is unknown. In this study live imaging of permissive and non-permissive cell lines, in conjunction with photo-bleaching (FRAP and FLIP), was used to investigate the trafficking of fluorescent labeled (EGFP) PRRSV-N protein. The data indicated that EGFP-PRRSV-N protein was not permanently sequestered to the nucleolus and had equivalent mobility to cellular nucleolar proteins. Further the nuclear import of N protein appeared to occur faster than nuclear export, which may account for the observed relative distribution of N protein between the cytoplasm and the nucleolus.

  7. Isolation and Sequence Analysis of Highly Pathogenic Porcine reproductive and respiratory syndrome virus from Swine Herds in the Jilin Province of China.

    PubMed

    Wang, Feng-Xue; Guo, Li; Yang, Yan-Ling; Song, Ni; Chen, Li-Zhi; Cheng, Shi-Peng; Wen, Yong-Jun

    2013-06-01

    The aim of the present study was to determine the causative agent of infected swines in the Jilin province of China and assess its genetic characteristics. Virus was isolated from tissues suspected of being infected by porcine reproductive and respiratory syndrome virus (PRRSV) and inoculated onto MARC-145 cells. Virus detection was carried out by RT-PCR, immunofluorescence, electron microscopy and sequencing. The results showed that the isolate was the North American genotype PRRSV, termed the JL-04/12 strain, with a 15,320 bp genome. The homology of the amino acid sequences in two nonstructural proteins and GP2 to GP5, between strains JL-04/12 and HUN4, ranged from 97.2 to 99.3 %. However, JL-04/12 GP6 and N protein were identical in HP-PRRSV JXA1 and HUN4. JL-04/12 was characterized by two discontinuous deletions in Nsp2. We speculate that the isolate is a variant of highly pathogenic porcine reproductive and respiratory syndrome derived from strains in 2006-2008. Altogether, these results indicate that highly pathogenic porcine reproductive and respiratory syndrome virus still exists in the Jilin province of China. PMID:24426266

  8. Effect of modified-live porcine reproductive and respiratory syndrome virus (PRRSv) vaccine on the shedding of wild-type virus from an infected population of growing pigs.

    PubMed

    Linhares, Daniel C L; Cano, Jean Paul; Wetzell, Thomas; Nerem, Joel; Torremorell, Montserrat; Dee, Scott A

    2012-01-01

    There are ongoing efforts to eliminate porcine reproductive and respiratory syndrome virus (PRRSv) from regions in the United States swine industry. However, an important challenge for the accomplishment of those efforts is the re-infection of pig units due to the area spread of PRRSv. The objective of this study was to evaluate the effect of PRRS modified-live virus vaccine (MLV) on viral shedding and on dynamics of PRRSv infection in pig populations raised under commercial conditions. The study composed of two rooms of 1000 pigs each. Ten percent of pigs of each room were inoculated with a field isolate of PRRSv. Rooms had separate air spaces and strict scientifically validated biosecurity protocols were adopted to avoid movement of pathogens between rooms. At 8 and 36 dpi (days post inoculation), all pigs of the challenge-vaccine group were inoculated with a MLV vaccine. Pigs of the challenge-control group were placebo-inoculated. Blood and oral fluid samples were collected from each room at 0, 8, 36, 70, 96 and 118 dpi for PRRSv RNA detection using PCR. PRRSv-antibodies were also screened from blood serum samples with a commercially available ELISA test. Additionally, tonsil scraping samples were collected from both groups at 70, 96 and 118 dpi. Moreover, air samples were collected 6 times per week from 0 to 118 dpi and were tested for PRRSv RNA using qPCR assay. There was no difference in the PRRSv infection dynamics measured as duration and magnitude of viremia and seroconversion. Also, there was no difference in the frequency of tonsil scraping samples PRRSv-positive by PCR. However, the challenge-vaccine group had significantly less PRRSv shed compared to the challenge-control group. The challenge-vaccine group had significant less PRRSv-positive oral fluids at 36 dpi. Moreover, the challenge-vaccine group had significant reduction in the cumulative PRRSv shed in the air. PMID:22063389

  9. The DNA virus white spot syndrome virus uses an internal ribosome entry site for translation of the highly expressed nonstructural protein ICP35.

    PubMed

    Kang, Shih-Ting; Wang, Han-Ching; Yang, Yi-Ting; Kou, Guang-Hsiung; Lo, Chu-Fang

    2013-12-01

    Although shrimp white spot syndrome virus (WSSV) is a large double-stranded DNA virus (?300 kbp), it expresses many polycistronic mRNAs that are likely to use internal ribosome entry site (IRES) elements for translation. A polycistronic mRNA encodes the gene of the highly expressed nonstructural protein ICP35, and here we use a dual-luciferase assay to demonstrate that this protein is translated cap independently by an IRES element located in the 5' untranslated region of icp35. A deletion analysis of this region showed that IRES activity was due to stem-loops VII and VIII. A promoterless assay, a reverse transcription-PCR together with quantitative real-time PCR analysis, and a stable stem-loop insertion upstream of the Renilla luciferase open reading frame were used, respectively, to rule out the possibility that cryptic promoter activity, abnormal splicing, or read-through was contributing to the IRES activity. In addition, a Northern blot analysis was used to confirm that only a single bicistronic mRNA was expressed. The importance of ICP35 to viral replication was demonstrated in a double-stranded RNA (dsRNA) interference knockdown experiment in which the mortality of the icp35 dsRNA group was significantly reduced. Tunicamycin was used to show that the ? subunit of eukaryotic initiation factor 2 is required for icp35 IRES activity. We also found that the intercalating drug quinacrine significantly inhibited icp35 IRES activity in vitro and reduced the mortality rate and viral copy number in WSSV-challenged shrimp. Lastly, in Sf9 insect cells, we found that knockdown of the gene for the Spodoptera frugiperda 40S ribosomal protein RPS10 decreased icp35 IRES-regulated firefly luciferase activity but had no effect on cap-dependent translation. PMID:24089551

  10. Association of the presence of influenza A virus and porcine reproductive and respiratory syndrome virus in sow farms with post-weaning mortality.

    PubMed

    Alvarez, Julio; Sarradell, Javier; Kerkaert, Barry; Bandyopadhyay, Dipankar; Torremorell, Montserrat; Morrison, Robert; Perez, Andres

    2015-10-01

    Influenza A virus (IAV) and porcine reproductive and respiratory syndrome virus (PRRSV) are among the most important pathogens affecting pigs worldwide. Their effect on post-weaning mortality can be substantial and may be potentiated by other concomitant factors. Here, the objective was to evaluate the association between IAV and PRRSV infection at weaning with post-weaning mortality observed in wean-to-finish farms in order to better quantify the full impact of their presence in breeding herds. IAV and PRRSV presence was assessed by real time reverse transcription (RRT)-PCR on oral fluid samples from suckling piglets in nine sow farms. Production data from 177 batches of growing pigs weaned one week before/after IAV and PRRSV testing were analyzed to measure the association between IAV and/or PRRSV test results and mortality recorded for a given batch through the use of Bayesian mixed effects negative binomial multivariable regression model. The model accounted for potential confounders such as flow, date at weaning, days on feed and batch size. A statistically important association between IAV (incidence ratio (IR)=1.18, 95% posterior probability interval 1.15-1.21) and PRRSV (IR=1.41, 95% PPI 1.30-1.52) with post-weaning mortality was detected, with season and number of days on feed also associated. Our results suggest that infection with IAV or PRRSV in the pre-weaning period is associated with an increase in post-weaning mortality. This association should be taken into consideration when measuring the impact of IAV and PRRSV in breeding herds. PMID:26210012

  11. Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water.

    PubMed

    Waiyapoka, Thanyaporn; Deachamag, Panchalika; Chotigeat, Wilaiwan; Bunsanong, Nittaya; Kanatharana, Proespichaya; Thavarungkul, Panote; Loyprasert-Thananimit, Suchera

    2015-10-01

    White spot syndrome virus (WSSV) is a major pathogen affecting the shrimp industry worldwide. In a preliminary study, WSSV binding protein (WBP) was specifically bound to the VP26 protein of WSSV. Therefore, we have developed the label-free affinity immunosensor using the WBP together with anti-GST-VP26 for quantitative detection of WSSV in shrimp pond water. When the biological molecules were immobilized on a gold electrode to form a self-assembled monolayer, it was then used to detect WSSV using a flow injection system with optimized conditions. Binding between the different copies of WSSV and the immobilized biological molecules was detected by an impedance change (?Z?) in real time. The sensitivity of the developed immunosensor was in the linear range of 1.6 × 10(1)-1.6 × 10(6) copies/?l. The system was highly sensitive for the analysis of WSSV as shown by the lack of impedance change when using yellow head virus (YHV). The developed immunosensor could be reused up to 37 times (relative standard deviation (RSD), 3.24 %) with a good reproducibility of residual activity (80-110 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation. PMID:26255303

  12. Cellular miR-130b inhibits replication of porcine reproductive and respiratory syndrome virus in vitro and in vivo.

    PubMed

    Li, Liwei; Gao, Fei; Jiang, Yifeng; Yu, Lingxue; Zhou, Yanjun; Zheng, Hao; Tong, Wu; Yang, Shen; Xia, Tianqi; Qu, Zehui; Tong, Guangzhi

    2015-01-01

    MicroRNAs (miRNAs) can impact viral infections by binding to sequences with partial complementarity on viral RNA transcripts, usually resulting in the repression of virus replication. In the present study, we identified a potential binding site for miR-130 in the 5' untranslated region (bps 155-162) of the porcine reproductive and respiratory syndrome virus (PRRSV) genome. We found that the delivery of multiple miR-130 family mimics, especially miR-130b, resulted in inhibition of PRRSV replication in vitro. miR-130 was effective in inhibiting the replication of multiple type 2 PRRSV strains, but not against vSHE, a classical type 1 strain. miR-130 over-expression did not induce IFN-? or TNF-? expression in either uninfected or PRRSV-infected porcine alveolar macrophages. Results from luciferase reporter assays indicated that miR-130 directly targeted the PRRSV 5' UTR. Intranasal inoculation of piglets with miR-130b exhibited antiviral activity in vivo and partially protected piglets from an otherwise lethal challenge with HP-PRRSV strain vJX143. Overall, these results demonstrate the importance of the miR-130 family in modulating PRRSV replication and also provide a scientific basis for using cellular miRNAs in anti-PRRSV therapies. PMID:26581169

  13. Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp

    SciTech Connect

    Liu Wangjing; Chang Yunshiang; Wang Chunghsiung; Kou, Guang-Hsiung; Lo Chufang . E-mail: gracelow@ntu.edu.tw

    2005-04-10

    Here, we report for the first time the successful use of cycloheximide (CHX) as an inhibitor to block de novo viral protein synthesis during WSSV (white spot syndrome virus) infection. Sixty candidate IE (immediate-early) genes were identified using a global analysis microarray technique. RT-PCR showed that the genes corresponding to ORF126, ORF242 and ORF418 in the Taiwan isolate were consistently CHX-insensitive, and these genes were designated ie1, ie2 and ie3, respectively. The sequences for these IE genes also appear in the two other WSSV isolates that have been sequenced. Three corresponding ORFs were identified in the China WSSV isolate, but only an ORF corresponding to ie1 was predicted in the Thailand isolate. In a promoter activity assay in Sf9 insect cells using EGFP (enhanced green fluorescence protein) as a reporter, ie1 showed very strong promoter activity, producing higher EGFP signals than the insect Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV) ie2 promoter.

  14. Cellular miR-130b inhibits replication of porcine reproductive and respiratory syndrome virus in vitro and in vivo

    PubMed Central

    Li, Liwei; Gao, Fei; Jiang, Yifeng; Yu, Lingxue; Zhou, Yanjun; Zheng, Hao; Tong, Wu; Yang, Shen; Xia, Tianqi; Qu, Zehui; Tong, Guangzhi

    2015-01-01

    MicroRNAs (miRNAs) can impact viral infections by binding to sequences with partial complementarity on viral RNA transcripts, usually resulting in the repression of virus replication. In the present study, we identified a potential binding site for miR-130 in the 5? untranslated region (bps 155-162) of the porcine reproductive and respiratory syndrome virus (PRRSV) genome. We found that the delivery of multiple miR-130 family mimics, especially miR-130b, resulted in inhibition of PRRSV replication in vitro. miR-130 was effective in inhibiting the replication of multiple type 2 PRRSV strains, but not against vSHE, a classical type 1 strain. miR-130 over-expression did not induce IFN-? or TNF-? expression in either uninfected or PRRSV-infected porcine alveolar macrophages. Results from luciferase reporter assays indicated that miR-130 directly targeted the PRRSV 5? UTR. Intranasal inoculation of piglets with miR-130b exhibited antiviral activity in vivo and partially protected piglets from an otherwise lethal challenge with HP-PRRSV strain vJX143. Overall, these results demonstrate the importance of the miR-130 family in modulating PRRSV replication and also provide a scientific basis for using cellular miRNAs in anti-PRRSV therapies. PMID:26581169

  15. Infectiousness of pigs infected by the Porcine Reproductive and Respiratory Syndrome virus (PRRSV) is time-dependent.

    PubMed

    Charpin, Céline; Mahé, Sophie; Keranflec'h, André; Belloc, Catherine; Cariolet, Roland; Le Potier, Marie-Frédérique; Rose, Nicolas

    2012-01-01

    The time-dependent transmission rate of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and the correlation between infectiousness, virological parameters and antibody responses of the infected pigs were studied in experimental conditions. Seven successive transmission trials involving a total of 77 specific pathogen-free piglets were carried out from 7 to 63 days post-inoculation (dpi). A semi-quantitative real time RT-PCR was developed to assess the evolution of the viral genome load in blood and nasal swabs from inoculated and contact pigs, with time. Virus genome in blood was detectable in inoculated pigs from 7 to 77 dpi, whereas viral genome shedding was detectable from nasal swabs from 2 to 48 dpi. The infectiousness of inoculated pigs, assessed from the frequency of occurrence of infected pigs in susceptible groups in each contact trial, increased from 7 to 14 dpi and then decreased slowly until 42 dpi (3, 7, 2, 1 and 0 pigs infected at 7, 14, 21, 28 and 42 dpi, respectively). These data were used to model the time-dependent infectiousness by a lognormal-like function with a latency period of 1 day and led to an estimated basic reproduction ratio, R0 of 2.6 [1.8, 3.3]. The evolution of infectiousness was mainly correlated with the time-course of viral genome load in the blood whereas the decrease of infectiousness was strongly related to the increase in total antibodies. PMID:23061672

  16. Efficient inhibition of porcine reproductive and respiratory syndrome virus replication by artificial microRNAs targeting the untranslated regions.

    PubMed

    Xia, Bing; Song, Hongqin; Chen, Yang; Zhang, Xinyu; Xia, Xiaoli; Sun, Huaichang

    2013-01-01

    A robust artificial microRNA (amiRNA) strategy against porcine reproductive and respiratory syndrome virus (PRRSV) was developed by targeting the untranslated regions (UTRs). Six candidate amiRNAs targeting the 5' or 3' UTR were used for vector construction, and four effective amiRNAs were selected for further study using a vector transfection/virus infection assay. In cell cultures stably transfected with the four amiRNA vectors, expression of the sequence-specific amiRNAs was confirmed using poly(A)-tailed RT-PCR. After infection with three different PRRSV strains, the viral RNA genome and/or transcript were inhibited by ~90 % (semi-quantitative RT-PCR), and the viral titers were decreased by more than six log CCID(50) (viral titration assay) before day 3 postinfection. The potent anti-PRRSV effects lasted for at least 5 days. Sequence analysis showed that the amiRNA antiviral activities were not compromised by the presence of one or two mismatches in their binding targets. This work constitutes a step towards developing a more effective RNAi strategy against PRRSV. PMID:22948796

  17. Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Inhibits RNA-Mediated Gene Silencing by Targeting Ago-2

    PubMed Central

    Chen, Jing; Shi, Xibao; Zhang, Xiaozhuan; Wang, Li; Luo, Jun; Xing, Guangxu; Deng, Ruiguang; Yang, Hong; Li, Jinting; Wang, Aiping; Zhang, Gaiping

    2015-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) infection strongly modulates the host’s immune response. The RNA silencing pathway is an intracellular innate response to viral infections. However, it is unknown whether PRRSV interacts with cellular RNA silencing to facilitate the viral infection. Here, we report for the first time the interaction between PRRSV and RNA silencing in both the porcine macrophages and African green monkey kidney cell line (MARC-145) cell line, which were derived from African green monkey kidney cells and highly permissive for PRRSV infection. Our data demonstrated that PRRSV suppressed RNA silencing induced by short-hairpin (sh) RNA, double-strand (ds) RNA and microRNA (miRNA) and downregulated the expression of argonaute protein-2 (Ago-2), which is a key protein of the RNA silencing pathway in animal cells. Further, exogenous introduction of siRNA and shRNA downregulated Dicer or Ago-2 proteins of the cellular RNA silencing apparatus in MARC-145 cells and porcine macrophages, which, in turn, increased the viral replication and titers. The viral non-structure protein 1? (nsp-1?) and nsp11 of PRRSV were identified as the suppressors for cellular RNA silencing (RSSs) to downregulate the Ago-2 protein. Our results identify that PRRSV, through its nsp proteins, suppresses the cellular RNA silencing apparatus in favor of viral infection and supports a co-evolutionary process of the virus and the cellular RNA silencing process. PMID:26512690

  18. Blood antioxidant enzymes (SOD, GPX), biochemical and haematological parameters in pigs naturally infected with porcine reproductive and respiratory syndrome virus.

    PubMed

    Stukelj, M; Toplak, I; Svete, A Nemec

    2013-01-01

    Porcine reproductive and respiratory syndrome (PRRS) has become one of the most economically important diseases for the swine industry worldwide. The objective of the study was to determine selected blood antioxidant enzymes (glutathione peroxidase (GPX), superoxide dismutase (SOD)), biochemical and haematological parameters in PRRS positive and negative pigs of three different categories, mainly to test oxidative stress hypothesis in pigs naturally infected with PRRS virus. Ninety PRRS positive and 90 PRRS negative pigs were included in the study. The presence of PRRS was confirmed by serological detection of antibodies against PRRS virus (PRRSV) and detection of PRRS viral RNA by RT-PCR. Pigs were further divided into three groups of 30: piglets just before weaning (weaners), fatteners and finishers. Blood samples for determining selected blood parameters were collected from the vena cava cranialis. Significantly (P < 0.05) higher activities of SOD in weaners and fatteners and of GPX in weaners were determined in PRRS positive pigs than in corresponding groups of PRRS negative pigs. In contrast, significantly (P < 0.05) lower GPX activity was observed in finishers of PRRS positive pigs than in the corresponding group of PRRS negative pigs. Concentrations of serum total protein in PRRS positive weaners and fatteners were significantly (P < 0.05) higher than those found in PRRS negative pigs. Leukopenia was observed in all three groups of PRRS positive pigs. It has been demonstrated, for the first time, that oxidative stress might be increased in PRRSV naturally infected pigs, especially in weaners. PMID:23971206

  19. Association between the genetic similarity of the open reading frame 5 sequence of Porcine reproductive and respiratory syndrome virus and the similarity in clinical signs of Porcine reproductive and respiratory syndrome in Ontario swine herds

    PubMed Central

    Rosendal, Thomas; Dewey, Cate; Friendship, Robert; Wootton, Sarah; Young, Beth; Poljak, Zvonimir

    2014-01-01

    A study of Ontario swine farms positive for Porcine reproductive and respiratory syndrome virus (PRRSV) tested the association between genetic similarity of the virus and similarity of clinical signs reported by the herd owner. Herds were included if a positive result of polymerase chain reaction for PRRSV at the Animal Health Laboratory at the University of Guelph, Guelph, Ontario, was found between September 2004 and August 2007. Nucleotide-sequence similarity and clinical similarity, as determined from a telephone survey, were calculated for all pairs of herds. The Mantel test indicated that clinical similarity and sequence similarity were weakly correlated for most clinical signs. The generalized additive model indicated that virus homology with 2 vaccine viruses affected the association between sequence similarity and clinical similarity. When the data for herds with vaccine-like virus were removed from the dataset there was a significant association between virus similarity and similarity of the reported presence of abortion, stillbirth, preweaning mortality, and sow/boar mortality. Ownership similarity was also found to be associated with virus similarity and with similarity of the reported presence of sows being off-feed, nursery respiratory disease, nursery mortality, finisher respiratory disease, and finisher mortality. These results indicate that clinical signs of PRRS are associated with PRRSV genotype and that herd ownership is associated with both of these. PMID:25355993

  20. Association between the genetic similarity of the open reading frame 5 sequence of Porcine reproductive and respiratory syndrome virus and the similarity in clinical signs of Porcine reproductive and respiratory syndrome in Ontario swine herds.

    PubMed

    Rosendal, Thomas; Dewey, Cate; Friendship, Robert; Wootton, Sarah; Young, Beth; Poljak, Zvonimir

    2014-10-01

    A study of Ontario swine farms positive for Porcine reproductive and respiratory syndrome virus (PRRSV) tested the association between genetic similarity of the virus and similarity of clinical signs reported by the herd owner. Herds were included if a positive result of polymerase chain reaction for PRRSV at the Animal Health Laboratory at the University of Guelph, Guelph, Ontario, was found between September 2004 and August 2007. Nucleotide-sequence similarity and clinical similarity, as determined from a telephone survey, were calculated for all pairs of herds. The Mantel test indicated that clinical similarity and sequence similarity were weakly correlated for most clinical signs. The generalized additive model indicated that virus homology with 2 vaccine viruses affected the association between sequence similarity and clinical similarity. When the data for herds with vaccine-like virus were removed from the dataset there was a significant association between virus similarity and similarity of the reported presence of abortion, stillbirth, preweaning mortality, and sow/boar mortality. Ownership similarity was also found to be associated with virus similarity and with similarity of the reported presence of sows being off-feed, nursery respiratory disease, nursery mortality, finisher respiratory disease, and finisher mortality. These results indicate that clinical signs of PRRS are associated with PRRSV genotype and that herd ownership is associated with both of these. PMID:25355993

  1. Modeling knowledge about possession transfer

    E-print Network

    Saliba, Gaylee (Gaylee Fouad)

    2009-01-01

    If we are to successfully create intelligent machines, it is essential to learn how to ground abstract notions, such as possession, in the physical world. In this work, I develop a model for the knowledge about possession ...

  2. Heartland Virus

    MedlinePLUS

    ... through the bite of a mosquito, tick, or sandfly. How do people get infected with Heartland virus? ... in the same genus as Rift Valley fever, Sandfly fever, Toscana, and Severe fever with thrombocytopenia syndrome ( ...

  3. Resolution of the cellular proteome of the nucleocapsid protein from a highly pathogenic isolate of porcine reproductive and respiratory syndrome virus identifies PARP-1 as a cellular target whose interaction is critical for virus biology

    PubMed Central

    Liu, Long; Lear, Zoe; Hughes, David J.; Wu, Weining; Zhou, En-min; Whitehouse, Adrian; Chen, Hongying; Hiscox, Julian A.

    2015-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat to the swine industry and food security worldwide. The nucleocapsid (N) protein is a major structural protein of PRRSV. The primary function of this protein is to encapsidate the viral RNA genome, and it is also thought to participate in the modulation of host cell biology and recruitment of cellular factors to facilitate virus infection. In order to the better understand these latter roles the cellular interactome of PRRSV N protein was defined using label free quantitative proteomics. This identified several cellular factors that could interact with the N protein including poly [ADP-ribose] polymerase 1 (PARP-1), a cellular protein, which can add adenosine diphosphate ribose to a protein. Use of the PARP-1 small molecule inhibitor, 3-AB, in PRRSV infected cells demonstrated that PARP-1 was required and acted as an enhancer factor for virus biology. Serial growth of PRRSV in different concentrations of 3-AB did not yield viruses that were able to grow with wild type kinetics, suggesting that by targeting a cellular protein crucial for virus biology, resistant phenotypes did not emerge. This study provides further evidence that cellular proteins, which are critical for virus biology, can also be targeted to ablate virus growth and provide a high barrier for the emergence of drug resistance. PMID:25614100

  4. Safety of Porcine Reproductive and Respiratory Syndrome Modified Live Virus (MLV) vaccine strains in a young pig infection model

    PubMed Central

    2013-01-01

    The objective of this study was to compare the safety of all modified live virus vaccines commercially available in Europe against Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) under the same experimental conditions. For this purpose, one hundred and twenty three-week-old piglets, divided into five groups, were used. On day 0 of the experiment, nine pigs per group were removed and the remaining fifteen were vaccinated with the commercial vaccines Ingelvac PRRS MLV, Amervac PRRS, Pyrsvac-183 and Porcilis PRRS by the IM route or were mock vaccinated and used as controls. On day 3, the nine unvaccinated pigs were re-introduced into their respective groups and served as sentinel pigs. Clinical signs were recorded daily and lung lesions were determined on days 7, 14 and 21, when 5 vaccinated pigs per group were euthanized. Blood samples and swabs were taken every three days and different organs were collected at necropsy to determine the presence of PRRSV. None of the vaccines studied caused detectable clinical signs in vaccinated pigs although lung lesions were found. Altogether, these results indicate that all vaccines can be considered clinically safe. However, some differences were found in virological parameters. Thus, neither Pyrsvac-183 nor Porcilis PRRS could be detected in porcine alveolar macrophage (PAM) cultures or in lung sections used to determine PRRSV by immunohistochemistry, indicating that these viruses might have lost their ability to replicate in PAM. This inability to replicate in PAM might be related to the lower transmission rate and the delay in the onset of viremia observed in these groups PMID:24308693

  5. Virulence and genotype-associated infectivity of interferon-treated macrophages by porcine reproductive and respiratory syndrome viruses.

    PubMed

    García-Nicolás, Obdulio; Baumann, Arnaud; Vielle, Nathalie Jane; Gómez-Laguna, Jaime; Quereda, Juan José; Pallarés, Francisco José; Ramis, Guillermo; Carrasco, Librado; Summerfield, Artur

    2014-01-22

    The polarization into M1 and M2 macrophages (M?) is essential to understand M? function. Consequently, the aim of this study was to determine the impact of IFN-? (M1), IL-4 (M2) and IFN-? activation of M? on the susceptibility to genotype 1 and 2 porcine reproductive respiratory syndrome (PRRS) virus (PRRSV) strains varying in virulence. To this end, monocyte-derived M? were generated by culture during 72h and polarization was induced for another 24h by addition of IFN-?, IL-4 or IFN-?. M? were infected with a collection of PRRSV isolates belonging to genotype 1 and genotype 2. Undifferentiated and M2 M? were highly susceptible to all PRRSV isolates. In contrast, M1 and IFN-? activated M? were resistant to low pathogenic genotype 1 PRRSV but not or only partially to genotype 2 PRRSV strains. Interestingly, highly virulent PRRSV isolates of both genotypes showed particularly high levels of infection compared with the prototype viruses in both M1 and IFN-?-treated M? (P<0.05). This was seen at the level of nucleocapsid expression, viral titres and virus-induced cell death. In conclusion, by using IFN-? and IFN-? stimulated M? it is possible to discriminate between PRRSV varying in genotype and virulence. Genotype 2 PRRSV strains are more efficient at escaping the intrinsic antiviral effects induced by type I and II IFNs. Our in vitro model will help to identify viral genetic elements responsible for virulence, an information important not only to understand PRRS pathogenesis but also for a rational vaccine design. Our results also suggest that monocyte-derived M? can be used as a PRRSV infection model instead of alveolar M?, avoiding the killing of pigs. PMID:24220223

  6. Induction of T helper 3 regulatory cells by dendritic cells infected with porcine reproductive and respiratory syndrome virus

    SciTech Connect

    Silva-Campa, Erika; Flores-Mendoza, Lilian; Resendiz, Monica; Pinelli-Saavedra, Araceli; Mata-Haro, Veronica; Mwangi, Waithaka; Hernandez, Jesus

    2009-05-10

    Delayed development of virus-specific immune response has been observed in pigs infected with the porcine reproductive and respiratory syndrome virus (PRRSV). Several studies support the hypothesis that the PRRSV is capable of modulating porcine immune system, but the mechanisms involved are yet to be defined. In this study, we evaluated the induction of T regulatory cells by PRRSV-infected dendritic cells (DCs). Our results showed that PRRSV-infected DCs significantly increased Foxp3{sup +}CD25{sup +} T cells, an effect that was reversible by IFN-alpha treatment, and this outcome was reproducible using two distinct PRRSV strains. Analysis of the expressed cytokines suggested that the induction of Foxp3{sup +}CD25{sup +} T cells is dependent on TGF-beta but not IL-10. In addition, a significant up-regulation of Foxp3 mRNA, but not TBX21 or GATA3, was detected. Importantly, our results showed that the induced Foxp3{sup +}CD25{sup +} T cells were able to suppress the proliferation of PHA-stimulated PBMCs. The T cells induced by the PRRSV-infected DCs fit the Foxp3{sup +}CD25{sup +} T helper 3 (Th3) regulatory cell phenotype described in the literature. The induction of this cell phenotype depended, at least in part, on PRRSV viability because IFN-alpha treatment or virus inactivation reversed these effects. In conclusion, this data supports the hypothesis that the PRRSV succeeds to establish and replicate in porcine cells early post-infection, in part, by inducing Th3 regulatory cells as a mechanism of modulating the porcine immune system.

  7. 'Kwanzan Stunting' syndrome: detection and molecular characterization of an Italian isolate of Little cherry virus 1.

    PubMed

    Matic, Slavica; Minafra, Angelantonio; Sánchez-Navarro, Jesús A; Pallás, Vicente; Myrta, Arben; Martelli, Giovanni P

    2009-07-01

    Evident stunting was observed for the first time on Prunus serrulata 'Kwanzan' indicator trees in Southern Italy during the indexing of two sour cherry accessions from cultivars 'Marasca di Verona' and 'Spanska'. Bud break and shooting were delayed and the developing leaves remained small. During the third year many Kwanzan plants died, regardless of the indexed cultivar. Electrophoretic analysis showed the presence of dsRNA pattern in extracts of stunted Kwanzan with a similar size to that of viruses of the family Closteroviridae. An identical pattern of more abundant dsRNA bands was obtained from GF305 seedlings grafted with the same sour cherry accessions. Observations by electron microscopy revealed the presence of long flexuous virus particles in both indicators (Kwanzan and GF305), characteristic of closteroviruses. Subsequent cloning work, starting from the dsRNA extracts of cultivar Marasca di Verona grafted on GF305 indicator, yielded 7 different clones, all showing high identity to the Little cherry virus 1 genome. Full sequencing of this virus isolate (ITMAR) was then done resulting in a complete genome composed of 16,936nt. Primers designed on the obtained sequences for RT-PCR detection confirmed the presence of Little cherry virus 1 in Kwanzan and GF305 trees, inoculated with both sour cherry cultivars. Phylogenetic analysis of the minor coat protein grouped virus isolates into two clusters: one including Italian isolates of sweet cherry, Japanese plum, peach and almond, together with German sweet cherry UW1 isolate, and a second one containing the Italian isolates of sour cherry (ITMAR and ITSPA), that were found associated with strong symptoms of 'Kwanzan Stunting'. PMID:19463722

  8. One case of swine hepatitis E virus and porcine reproductive and respiratory syndrome virus Co-infection in weaned pigs

    PubMed Central

    2013-01-01

    Background Using various methods, we analyzed the cause of death among weaned pigs from a pig farm in Hebei Province, China. All 300 piglets (100% fatality) were identified as moribund, with death occurring within 1 month from the onset of clinical signs. Results A single case exhibited obvious hemorrhagic necrotic changes with massive lymphocytic infiltration in multiple organs, in particular the liver, lungs and intestines. Dysplasia and lymphocyte deterioration were common in lymphatic organs. No visible bacterial colonies from liver and spleen were observed in nutrient, MacConkey, and blood agar plates. Using polymerase chain reaction techniques for this case, we attempted to detect a number of epidemic swine viruses in spleen and liver, including PRRSV, CSF, HEV, and PCV2. We found that this sample was positive for the presence of HEV and PRRSV. Conclusions We have detected HEV and PRRSV co-infection in one piglet. Severe pathologic changes were observed. The high mortality of weaned pigs which showed the similar clinical syptom was possibly a result of HEV and PRRSV co-infection, which has rarely been reported previously. We speculated that co-infection with PRRSV and HEV might lead to more serious problems. PMID:24252365

  9. Multicentric plasmocytic Castleman's disease with polyneuropathy, organomegaly, endocrinopathy, M protein, skin changes syndrome and coexistent human herpes virus-6 infection--a possible relationship.

    PubMed

    Fazakas, Adám; Csire, Márta; Berencsi, György; Szepesi, Agota; Matolcsy, András; Jakab, Lajos; Karádi, István; Várkonyi, Judit

    2009-10-01

    Authors report a case of Castleman's disease (CD) with polyneuropathy, organomegaly, endocrinopathy, M protein, skin change (POEMS) syndrome. According to the present knowledge, these two rare conditions are often induced by Human Herpes Vírus- 8 (HHV-8) or by Human Immunodefeciency Virus, separately or in combination. In this case, however, HHV-6 viral DNA had been detected in the blood and lymph node samples by PCR. The authors conclude that the modulation of immune functions by HHV-6 might be responsible for the development of CD and POEMS syndrome in the referred case. PMID:19863339

  10. UNDERSTANDING SWINE IMMUNITY TO PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS (PRRSV) INFECTION - INFORMING FUTURE VACCINE DESIGN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Porcine Reproductive and Respiratory Syndrome (PRRS) is the most economically significant disease facing the swine industry today, costing U.S. pork producers at least $560 million annually. This abstract describes some of the approaches we’ve tested to evaluate immunity to PRRSV. We plan to use th...

  11. In Vivo Growth of Porcine Reproductive and Respiratory Syndrome Virus Engineered Nsp2 Deletion Mutants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Prior studies on PRRSV strain VR-2332 nonstructural protein 2 (nsp2) had shown that as much as 403 amino acids could be removed from the hypervariable region without losing virus viability in vitro. We utilized selected nsp2 deletion mutants to examine in vivo growth. Young swine (4 pigs/group; 5 co...

  12. Innate and adaptive immunity against porcine reproductive and respiratory syndrome virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many highly effective vaccines have been produced against viruses whose virulent infection elicits strong and durable protective immunity. In these cases, characterization of immune effector mechanisms and identification of protective epitopes/immunogens has been informative for the development of s...

  13. Rooting the Phylogenetic Tree of Middle East Respiratory Syndrome Coronavirus by Characterization of a Conspecific Virus from an African Bat

    PubMed Central

    Corman, Victor Max; Ithete, Ndapewa Laudika; Richards, Leigh Rosanne; Schoeman, M. Corrie; Preiser, Wolfgang

    2014-01-01

    ABSTRACT The emerging Middle East respiratory syndrome coronavirus (MERS-CoV) causes lethal respiratory infections mainly on the Arabian Peninsula. The evolutionary origins of MERS-CoV are unknown. We determined the full genome sequence of a CoV directly from fecal material obtained from a South African Neoromicia capensis bat (NeoCoV). NeoCoV shared essential details of genome architecture with MERS-CoV. Eighty-five percent of the NeoCoV genome was identical to MERS-CoV at the nucleotide level. Based on taxonomic criteria, NeoCoV and MERS-CoV belonged to one viral species. The presence of a genetically divergent S1 subunit within the NeoCoV spike gene indicated that intraspike recombination events may have been involved in the emergence of MERS-CoV. NeoCoV constitutes a sister taxon of MERS-CoV, placing the MERS-CoV root between a recently described virus from African camels and all other viruses. This suggests a higher level of viral diversity in camels than in humans. Together with serologic evidence for widespread MERS-CoV infection in camelids sampled up to 20 years ago in Africa and the Arabian Peninsula, the genetic data indicate that camels act as sources of virus for humans rather than vice versa. The majority of camels on the Arabian Peninsula is imported from the Greater Horn of Africa, where several Neoromicia species occur. The acquisition of MERS-CoV by camels from bats might have taken place in sub-Saharan Africa. Camelids may represent mixing vessels for MERS-CoV and other mammalian CoVs. IMPORTANCE It is unclear how, when, and where the highly pathogenic MERS-CoV emerged. We characterized the full genome of an African bat virus closely related to MERS-CoV and show that human, camel, and bat viruses belong to the same viral species. The bat virus roots the phylogenetic tree of MERS-CoV, providing evidence for an evolution of MERS-CoV in camels that preceded that in humans. The revised tree suggests that humans are infected by camels rather than vice versa. Although MERS-CoV cases occur mainly on the Arabian Peninsula, the data from this study together with serologic and molecular investigations of African camels indicate that the initial host switch from bats may have taken place in Africa. The emergence of MERS-CoV likely involved exchanges of genetic elements between different viral ancestors. These exchanges may have taken place either in bat ancestors or in camels acting as mixing vessels for viruses from different hosts. PMID:25031349

  14. Complete Genome Sequence of a Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Variant with a New Insertion in Glycoprotein 5, Isolated from a Stillborn Fetus

    PubMed Central

    Liu, Can; Zhang, Lei

    2015-01-01

    Here, we report the complete genome sequence of strain GD-2011, a highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) that was isolated from a stillborn fetus. The GD-2011 strain is characterized by a discontinuous 30–amino acid deletion in the nonstructural protein 2. In addition, GD-2011 had a 1–amino acid insertion in glycoprotein 5, which does not exist in any other HP-PRRSV strains. PMID:26607885

  15. Complete Genome Sequence of Type 1 Porcine Reproductive and Respiratory Syndrome Virus Strain E38, Isolated from South Korea with a Novel Deletion

    PubMed Central

    Kim, Jeong-Min; Kwon, Young-Woo; Choi, Eun-Jin; Ouh, In-Ohk; Choe, Se-Eun; Lee, Jienny; Song, Jae-Young

    2015-01-01

    We report the complete genome sequence of the European type 1 porcine reproductive and respiratory syndrome virus E38 strain, isolated from South Korea with a novel deletion. It contains a 61-nucleotide discontinuous deletion of the Nsp2 and Nsp12 regions. This study will aid in understanding the genetic diversity of type 1 PRRSV and in manufacturing a construct based on Korean vaccine candidate development. PMID:26472832

  16. The Neuraminidase Inhibitor Oseltamivir Is Effective Against A/Anhui/1/2013 (H7N9) Influenza Virus in a Mouse Model of Acute Respiratory Distress Syndrome

    PubMed Central

    Baranovich, Tatiana; Burnham, Andrew J.; Marathe, Bindumadhav M.; Armstrong, Jianling; Guan, Yi; Shu, Yuelong; Peiris, Joseph Malik Sriyal; Webby, Richard J.; Webster, Robert G.; Govorkova, Elena A.

    2014-01-01

    Background.?High mortality and uncertainty about the effectiveness of neuraminidase inhibitors (NAIs) in humans infected with influenza A(H7N9) viruses are public health concerns. Methods.?Susceptibility of N9 viruses to NAIs was determined in a fluorescence-based assay. The NAI oseltamivir (5, 20, or 80 mg/kg/day) was administered to BALB/c mice twice daily starting 24, 48, or 72 hours after A/Anhui/1/2013 (H7N9) virus challenge. Results.?All 12 avian N9 and 3 human H7N9 influenza viruses tested were susceptible to NAIs. Without prior adaptation, A/Anhui/1/2013 (H7N9) caused lethal infection in mice that was restricted to the respiratory tract and resulted in pulmonary edema and acute lung injury with hyaline membrane formation, leading to decreased oxygenation, all characteristics of human acute respiratory distress syndrome. Oseltamivir at 20 and 80 mg/kg protected 80% and 88% of mice when initiated after 24 hours, and the efficacy decreased to 70% and 60%, respectively, when treatment was delayed by 48 hours. Emergence of oseltamivir-resistant variants was not detected. Conclusions.?H7N9 viruses are comparable to currently circulating influenza A viruses in susceptibility to NAIs. Based on these animal studies, early treatment is associated with improved outcomes. PMID:24133191

  17. Probe-free real-time reverse transcription polymerase chain reaction assays for the detection and typing of porcine reproductive and respiratory syndrome virus in Canada

    PubMed Central

    Eschbaumer, Michael; Li, Wansi (May); Wernike, Kerstin; Marshall, Frank; Czub, Markus

    2015-01-01

    Porcine reproductive and respiratory syndrome (PRRS) has tremendous impact on the pork industry in North America. The molecular diagnosis of infection with PRRS virus (PRRSV) is hampered by its considerable strain diversity. In this study, 43 previously published or newly developed primers for probe-free real-time reverse transcription polymerase chain reaction (RT-PCR) were evaluated on their sensitivity, specificity, reproducibility, and repeatability, using a diverse panel of 36 PRRSV strains as well as other arteriviruses and unrelated porcine viruses. Three primer pairs had excellent diagnostic and analytical sensitivity on par with a probe-based reference assay, absolute specificity to virus genotype and species, as well as over 95% reproducibility and repeatability across a wide dynamic range. PMID:26130848

  18. Probe-free real-time reverse transcription polymerase chain reaction assays for the detection and typing of porcine reproductive and respiratory syndrome virus in Canada.

    PubMed

    Eschbaumer, Michael; Li, Wansi May; Wernike, Kerstin; Marshall, Frank; Czub, Markus

    2015-07-01

    Porcine reproductive and respiratory syndrome (PRRS) has tremendous impact on the pork industry in North America. The molecular diagnosis of infection with PRRS virus (PRRSV) is hampered by its considerable strain diversity. In this study, 43 previously published or newly developed primers for probe-free real-time reverse transcription polymerase chain reaction (RT-PCR) were evaluated on their sensitivity, specificity, reproducibility, and repeatability, using a diverse panel of 36 PRRSV strains as well as other arteriviruses and unrelated porcine viruses. Three primer pairs had excellent diagnostic and analytical sensitivity on par with a probe-based reference assay, absolute specificity to virus genotype and species, as well as over 95% reproducibility and repeatability across a wide dynamic range. PMID:26130848

  19. Structure of Severe Fever with Thrombocytopenia Syndrome Virus Nucleocapsid Protein in Complex with Suramin Reveals Therapeutic Potential

    PubMed Central

    Jiao, Lianying; Ouyang, Songying; Liang, Mifang; Niu, Fengfeng; Shaw, Neil; Wu, Wei; Ding, Wei; Jin, Cong; Peng, Yao; Zhu, Yanping; Zhang, Fushun; Wang, Tao; Li, Chuan; Zuo, Xiaobing; Luan, Chi-Hao; Li, Dexin

    2013-01-01

    Severe fever with thrombocytopenia syndrome is an emerging infectious disease caused by a novel bunyavirus (SFTSV). Lack of vaccines and inadequate therapeutic treatments have made the spread of the virus a global concern. Viral nucleocapsid protein (N) is essential for its transcription and replication. Here, we present the crystal structures of N from SFTSV and its homologs from Buenaventura (BUE) and Granada (GRA) viruses. The structures reveal that phleboviral N folds into a compact core domain and an extended N-terminal arm that mediates oligomerization, such as tetramer, pentamer, and hexamer of N assemblies. Structural superimposition indicates that phleboviral N adopts a conserved architecture and uses a similar RNA encapsidation strategy as that of RVFV-N. The RNA binding cavity runs along the inner edge of the ring-like assembly. A triple mutant of SFTSV-N, R64D/K67D/K74D, almost lost its ability to bind RNA in vitro, is deficient in its ability to transcribe and replicate. Structural studies of the mutant reveal that both alterations in quaternary assembly and the charge distribution contribute to the loss of RNA binding. In the screening of inhibitors Suramin was identified to bind phleboviral N specifically. The complex crystal structure of SFTSV-N with Suramin was refined to a 2.30-Å resolution. Suramin was found sitting in the putative RNA binding cavity of SFTSV-N. The inhibitory effect of Suramin on SFTSV replication was confirmed in Vero cells. Therefore, a common Suramin-based therapeutic approach targeting SFTSV-N and its homologs could be developed for containing phleboviral outbreaks. PMID:23576501

  20. RNA interference targeting nucleocapsid protein inhibits porcine reproductive and respiratory syndrome virus replication in Marc-145 cells.

    PubMed

    Yang, Minnan; Xiang, Qun; Zhang, Xiaodong; Li, Xiang; Sylla, Seydou; Ding, Zhuang

    2014-04-01

    Porcine reproductive and respiratory syndrome (PRRS) is an important disease, which leads to severe economic losses in swine-producing areas of the world. However, current antiviral strategies cannot provide highly effective protection. In this study, three theoretically effective interference target sites (71-91, 144-164, 218-238) targeting the nucleocapsid (N) gene of PRRSV were designed and selected, and then three siRNA-expressing plasmids were constructed, respectively named p2.1-N71, p2.1-N144, and p2.1-N218. The recombinant siRNA-expressing plasmids were transfected into Marc-145 cells; then the cells were infected with PRRSV (JL07SW strain); finally, after incubation for 48 h, the antiviral activity of those siRNA-expressing plasmids in Marc-145 cells was assessed by cytopathic effects, virus titers, indirect immunofluorescence, and quantitative real-time PCR. Experimental results demonstrated that these three siRNA-expressing plasmids could effectively and significantly inhibit the replication of PRRSV by 93.2%, 83.6%, and 89.2% in Marc-145 cells, respectively. Among these three siRNA-expressing plasmids, p2.1-N71 was found to be most effective, while p2.1-N144 and p2.1-N218 displayed relatively weak inhibition of virus replication. The results indicated that siRNA-expressing plasmids targeting the N gene of PRRSV could significantly inhibit PRRSV replication in Marc-145 cells. Based on our experimental results and previous reports, the 71-91, 179-197, and 234-252 sites of the N gene are good choices to effectively inhibit the replication of PRRSV, and this RNA interference technique can be a potential anti-PRRSV strategy. PMID:24682995

  1. Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon

    SciTech Connect

    Syed Musthaq, S.; Madhan, Selvaraj; Sahul Hameed, A.S.; Kwang, Jimmy

    2009-09-01

    White spot syndrome virus (WSSV) is a large dsDNA virus responsible for white spot disease in shrimp and other crustaceans. VP28 is one of the major envelope proteins of WSSV and plays a crucial role in viral infection. In an effort to develop a vaccine against WSSV, we have constructed a recombinant baculovirus with an immediate early promoter 1 which expresses VP28 at an early stage of infection in insect cells. Baculovirus expressed rVP28 was able to maintain its structural and antigenic conformity as indicated by immunofluorescence assay and western blot analysis. Interestingly, our results with confocal microscopy revealed that rVP28 was able to localize on the plasma membrane of insect cells infected with recombinant baculovirus. In addition, we demonstrated with transmission electron microscopy that baculovirus successfully acquired rVP28 from the insect cell membrane via the budding process. Using this baculovirus displaying VP28 as a vaccine against WSSV, we observed a significantly higher survival rate of 86.3% and 73.5% of WSSV-infected shrimp at 3 and 15 days post vaccination respectively. Quantitative real-time PCR also indicated that the WSSV viral load in vaccinated shrimp was significantly reduced at 7 days post challenge. Furthermore, our RT-PCR and immunohistochemistry results demonstrated that the recombinant baculovirus was able to express VP28 in vivo in shrimp tissues. This study will be of considerable significance in elucidating the morphogenesis of WSSV and will pave the way for new generation vaccines against WSSV.

  2. An intracellularly expressed Nsp9-specific nanobody in MARC-145 cells inhibits porcine reproductive and respiratory syndrome virus replication.

    PubMed

    Liu, Hongliang; Wang, Yan; Duan, Hong; Zhang, Angke; Liang, Chao; Gao, Jiming; Zhang, Chong; Huang, Baicheng; Li, Qiongyi; Li, Na; Xiao, Shuqi; Zhou, En-Min

    2015-12-31

    Porcine reproductive and respiratory syndrome (PRRS) is a widespread viral disease affecting the swine industry, with no cure or effective treatment. Current vaccines are inefficient mainly due to the high degree of genetic and antigenic variation within PRRS virus (PRRSV) strains. Thus, the development of novel anti-PRRSV strategies is an important area of research. The nonstructural protein 9 (Nsp9) of PRRSV is essential for viral replication, and its sequence is relatively conserved, making it a logical antiviral target for PRRSV. Camel single-domain antibodies (nanobodies) represent a promising antiviral approach because of their small size, high specificity, and solubility. However, no nanobodies against PRRSV have been reported to date. In this study, Nsp9-specific nanobodies were isolated from a phage display library of variable domains of Camellidaeheavy chain-only antibodies (VHH). One of the isolated nanobodies, Nb6, was chosen for further investigation. Co-immunoprecipitation experiments indicated that Nb6 can still maintain antigen binding capabilities when expressed in the cell cytoplasm. A MARC-145 cell line stably expressing Nb6 was established to investigate its potential antiviral activity. Our results showed that intracellularly expressed Nb6 could potently suppress PRRSV replication by inhibiting viral genome replication and transcription. More importantly, Nb6 could protect MARC-145 cells from virus-induced cytopathic effect (CPE) and fully block PRRSV replication at an MOI of 0.01 or lower. To our knowledge, this is the first report of a nanobody based antiviral strategy against PRRSV, and this finding has the potential to lead to future developments of novel antiviral treatments for PRRSV infection. PMID:26525739

  3. Pathogenesis of Korean type 1 (European genotype) porcine reproductive and respiratory syndrome virus in experimentally infected pigs.

    PubMed

    Han, K; Seo, H W; Oh, Y; Kang, I; Park, C; Chae, C

    2012-01-01

    The aim of this study was to elucidate the pathogenesis of experimental infection with Korean type 1 porcine reproductive and respiratory syndrome virus (PRRSV) by defining the virus distribution, sites of viral replication, viraemia and gross and microscopical lesions in conventional pigs studied for 28 days after intranasal inoculation. Mean rectal temperature was significantly higher in infected pigs than in negative control pigs at 2 days post inoculation (dpi) (P=0.004), 3 dpi (P<0.001), 4 dpi (P=0.003) and 5 dpi (P=0.034). The log(10)TCID(50)/ml of type 1 PRRSV increased significantly at 0-1 dpi (P=0.024) and 5-7 dpi (P=0.029), but decreased at 10-14 dpi (P=0.026) and 14-21 dpi (P=0.012) in infected pigs. Infected pigs developed multifocal, tan-mottled areas of lung tissue with irregular and indistinct borders. Microscopical lesions, when present, were multifocal, mild to moderate, generally most extensive at 5-7 dpi (P=0.036), and were nearly resolved at 28 dpi. Type 1 PRRSV nucleic acid and antigen were detected exclusively within the cytoplasm of macrophages and type I and II pneumocytes. The score for PRRSV-positive cells increased at 3-7 dpi (P<0.05) and decreased at 10-14 dpi (P=0.034) in infected pigs. Thus, respiratory disease was reproduced in conventional pigs by infection with Korean type 1 PRRSV. PMID:22316433

  4. Expression of the nucleocapsid protein of porcine reproductive and respiratory syndrome virus in soybean seed yields an immunogenic antigenic protein.

    PubMed

    Vimolmangkang, Sornkanok; Gasic, Ksenija; Soria-Guerra, Ruth; Rosales-Mendoza, Sergio; Moreno-Fierros, Leticia; Korban, Schuyler S

    2012-03-01

    Porcine reproductive and respiratory syndrome (PRRS), caused by the PRRS virus (PRRSV), is a serious disease of swine and contributes to severe worldwide economic losses in swine production. Current vaccines against PRRS rely on the use of an attenuated-live virus; however, these are unreliable. Thus, alternative effective vaccines against PRRS are needed. Plant-based subunit vaccines offer viable, safe, and environmentally friendly alternatives to conventional vaccines. In this study, efforts have been undertaken to develop a soybean-based vaccine against PRRSV. A construct carrying a synthesized PRRSV-ORF7 antigen, nucleocapsid N protein of PRRSV, has been introduced into soybean, Glycine max (L.) Merrill. cvs. Jack and Kunitz, using Agrobacterium-mediated transformation. Transgenic plants carrying the sORF7 transgene have been successfully generated. Molecular analyses of T(0) plants confirmed integration of the transgene and transcription of the PRRSV-ORF7. Presence of a 15-kDa protein in seeds of T(1) transgenic lines was confirmed by Western blot analysis using PRRSV-ORF7 antisera. The amount of the antigenic protein accumulating in seeds of these transgenic lines was up to 0.65% of the total soluble protein (TSP). A significant induction of a specific immune response, both humoral and mucosal, against PRRSV-ORF7 was observed following intragastric immunization of BALB/c female mice with transgenic soybean seeds. These findings provide a 'proof of concept', and serve as a critical step in the development of a subunit plant-based vaccine against PRRS. PMID:21971995

  5. Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT)

    PubMed Central

    Xie, Shijun; Zhang, Xiaojun; Zhang, Jiquan; Li, Fuhua; Xiang, Jianhai

    2015-01-01

    White spot syndrome virus (WSSV) is a major pathogen in shrimp cultures. The interactions between viral proteins and their receptors on the surface of cells in a frontier target tissue are crucial for triggering an infection. In this study, a yeast two-hybrid (Y2H) library was constructed using cDNA obtained from the stomach and gut of Litopenaeus vannamei, to ascertain the role of envelope proteins in WSSV infection. For this purpose, VP37 was used as the bait in the Y2H library screening. Forty positive clones were detected after screening. The positive clones were analyzed and discriminated, and two clones belonging to the peritrophin family were subsequently confirmed as genuine positive clones. Sequence analysis revealed that both clones could be considered as the same gene, LV-peritrophin (LvPT). Co-immunoprecipitation confirmed the interaction between LvPT and VP37. Further studies in the Y2H system revealed that LvPT could also interact with other WSSV envelope proteins such as VP32, VP38A, VP39B, and VP41A. The distribution of LvPT in tissues revealed that LvPT was mainly expressed in the stomach than in other tissues. In addition, LvPT was found to be a secretory protein, and its chitin-binding ability was also confirmed. PMID:26692362

  6. First detection of severe fever with thrombocytopenia syndrome virus in the tick species Haemaphysalis concinna in Shandong Province, China.

    PubMed

    Meng, Kai; Sun, Wenjing; Cheng, Ziqiang; Guo, Huijun; Liu, Jianzhu; Chai, Tongjie

    2015-12-01

    The aim of this study was to detect severe fever with thrombocytopenia syndrome virus (SFTSV) infection using polymerase chain reaction (PCR) amplification in adult Haemaphysalis concinna ticks. A total of 72 adult H. concinna ticks were obtained from 35 goats, three adult H. concinna ticks (4.17 %) collected from two goats were found to be infected with SFTSV via PCR assay. Sequence analysis showed that the partial segment M glycoprotein gene of SFTSV was about 500 bases long by polymerase chain reaction (PCR) amplification and that the PCR products from the samples had an identical sequence (KP714259). With regard to the phylogenetic analysis, the Nei-Gojobri (Kimura 2-parameter) method was used to construct the phylogenetic trees. Phylogenetic analysis indicated that the obtained sequence closely resembled SFTSV strain from Zhejiang Province (KC189856) and belonged to the same clade. The similarity of these strains was up to 96.62 % (only differing by 17 bases). In addition, phylogenetic analysis also indicated that the sequence obtained from adult H. concinna ticks was most closely related to the sequence isolated from Haemaphysalis longicornis (KF781498) with 97.22 % similarity (differing only by 4 bases) and belonged to the same clade. PMID:26350381

  7. Graphene oxide based fluorescence resonance energy transfer and loop-mediated isothermal amplification for white spot syndrome virus detection.

    PubMed

    Waiwijit, U; Phokaratkul, D; Kampeera, J; Lomas, T; Wisitsoraat, A; Kiatpathomchai, W; Tuantranont, A

    2015-10-20

    Graphene oxide (GO) is attractived for biological or medical applications due to its unique electrical, physical, optical and biological properties. In particular, GO can adsorb DNA via ?-? stacking or non-covalent interactions, leading to fluorescence quenching phenomenon applicable for bio-molecular detection. In this work, a new method for white spot syndrome virus (WSSV)-DNA detection is developed based on loop-mediated isothermal amplification (LAMP) combined with fluorescence resonance energy transfer (FRET) between GO and fluorescein isothiocyanate-labeled probe (FITC-probe). The fluorescence quenching efficiency of FITC-probe was found to increase with increasing GO concentration and reached 98.7% at a GO concentration of 50?g/ml. The fluorescence intensity of FITC-probe was recovered after hybridization with WSSV LAMP product with an optimal hybridization time of 10min and increased accordingly with increasing amount of LAMP products. The detection limit was estimated to be as low as 10 copies of WSSV plasmid DNA or 0.6fg of the total DNA extracted from shrimp infected with WSSV. In addition, no cross reaction was observed with other common shrimp viral pathogens. Therefore, the GO-FRET-LAMP technique is promising for fast, sensitive and specific detection of DNAs. PMID:26277651

  8. Efficacy of combined vaccination against Mycoplasma hyopneumoniae and porcine reproductive and respiratory syndrome virus in dually infected pigs.

    PubMed

    Bourry, Olivier; Fablet, Christelle; Simon, Gaëlle; Marois-Créhan, Corinne

    2015-11-18

    Porcine respiratory disease complex (PRDC) is one of the main causes of economic losses for swine producers. This complex is due to a combination of different pathogens and their interactions. Two major pathogens involved in PRDC are Mycoplasma hyopneumoniae (Mhp) and porcine reproductive and respiratory syndrome virus (PRRSV). The objectives of this study were (i) to develop an experimental model of dual Mhp/PRRSV infection in SPF pigs with European strains of Mhp and PRRSV and (ii) to assess and compare the effects of single Mhp, single PRRSV or combined Mhp/PRRSV vaccination against this dual infection. Pigs dually infected with Mhp and PRRSV showed a combination of symptoms characteristic of each pathogen but no significant exacerbation of pathogenicity. Thus, the co-infected pigs displayed coughing and pneumonia typical of Mhp infection in addition to PRRSV-related hyperthermia and decrease in average daily gain (ADG). Hyperthermia was reduced in PRRSV vaccinated animals (single or combined vaccination), whereas ADG was restored in Mhp/PRRSV vaccinated pigs only. Regarding respiratory symptoms and lung lesions, no vaccine decreased coughing. However, all vaccines reduced the pneumonia score but more so in animals receiving the Mhp vaccine, whether single or combined. This vaccine also decreased the Mhp load in the respiratory tract. In conclusion, combined vaccination against both Mhp and PRRSV efficiently pooled the efficacy of each single PRRSV and Mhp vaccination and could be an interesting tool to control PRDC in European swine production. PMID:26422712

  9. Quantitation of varicella-zoster virus DNA in patients with Ramsay Hunt syndrome and zoster sine herpete.

    PubMed

    Furuta, Y; Ohtani, F; Sawa, H; Fukuda, S; Inuyama, Y

    2001-08-01

    Varicella-zoster virus (VZV) reactivation causes facial nerve palsy in Ramsay Hunt syndrome (RHS) and zoster sine herpete (ZSH) with and without zoster rash, respectively. In the present study, we analyzed the VZV DNA copy number in saliva samples from 25 patients with RHS and 31 patients with ZSH using a TaqMan PCR assay to determine differences in the viral load between the two diseases. VZV copy number in saliva peaked near the day of the appearance of zoster in patients with RHS. Consequently, VZV DNA was less frequently detected in patients with RHS who exhibited facial palsy several days after the appearance of zoster. These findings suggest that the VZV load in saliva samples reflects the kinetics of viral reactivation in patients with RHS. In addition, VZV DNA was equally detected in saliva from patients with RHS and ZSH, and there was no significant difference in the highest viral copy number between patients with RHS and those with ZSH. The VZV load does not appear to reflect a major difference between RHS and ZSH. PMID:11474003

  10. Cytokines transcript levels in lung and lymphoid organs during genotype 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection.

    PubMed

    García-Nicolás, Obdulio; Quereda, Juan José; Gómez-Laguna, Jaime; Salguero, Francisco Javier; Carrasco, Librado; Ramis, Guillermo; Pallarés, Francisco José

    2014-07-15

    Porcine Reproductive and Respiratory Syndrome (PRRS) is one of the most economically important diseases of swine. PRRS virus (PRRSV) infection in the pig is characterized by a weak or absent host innate immune response. The underlying mechanisms of PRRSV pathogenesis are still unclear. The analysis of transcript levels represents an alternative to immunoassays for the detection of cytokines that sometimes are difficult to detect due to their low amounts. This study sets out to determine the differences in pathogenesis and the immune response between lung, tonsil, tracheobronchial lymph node (Tb-LN) and retropharyngeal LN (Rf-LN) of PRRSV 2982 strain infected pigs. PRRSV strain 2982 avoided the onset of an effective innate immune response, especially in PRRSV main target (lung) and reservoir (tonsil) organs. PRRSV lead to an impaired expression of IFN-? and TNF-? gene expression, which finally induced a weak and delayed adaptive immune response through an inefficient IL-12 and IFN-? expression. Finally, PRRSV replication favored the expression of the anti-inflammatory IL-10 cytokine in infected pigs. PMID:24726859

  11. Rapid Tests and the Diagnosis of Visceral Leishmaniasis and Human Immunodeficiency Virus/Acquired Immunodeficiency Syndrome Coinfection.

    PubMed

    Barbosa Júnior, Walter Lins; Ramos de Araújo, Paulo Sérgio; Dias de Andrade, Luiz; Aguiar Dos Santos, Ana Maria; Lopes da Silva, Maria Almerice; Dantas-Torres, Filipe; Medeiros, Zulma

    2015-11-01

    After the emergence of the human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS), the number of visceral leishmaniasis (VL)-HIV/AIDS coinfections has increased worldwide. Herein, we assessed the usefulness of an rK39-based immunochromatographic test (rK39 ICT) (DiaMed-IT LEISH(®); DiaMed AG, Cressier-sur-Morat, Switzerland) and a latex agglutination test (KAtex; Kalon Biological, Guildford, United Kingdom) for urinary antigen detection to diagnose VL in 15 HIV/AIDS patients from northeastern Brazil. VL diagnosis was based on clinical findings, cytology, serology, parasite DNA, and/or urinary antigen detection. VL was confirmed in seven out of 15 HIV/AIDS patients. Only three patients were positive in bone marrow cytology, three patients were conventional polymerase chain reaction (PCR) positive, while six were real-time PCR positive. All patients were direct agglutination test (DAT) (Royal Tropical Institute, Amsterdam, The Netherlands) positive; of these, four were positive by rK39 ICT and five by KAtex. Large-scale studies are needed to validate the use of the KAtex in the national public health laboratory network in Brazil, aiming at improving the diagnosis of VL in HIV/AIDS patients in this country. PMID:26416105

  12. Label Free Detection of White Spot Syndrome Virus Using Lead Magnesium Niobate-Lead Titanate Piezoelectric Microcantilever Sensors

    PubMed Central

    Capobianco, Joseph; Shih, Wei-Heng; Leu, Jiann-Horng; Lo, Grace Chu-Fang; Shih, Wan Y.

    2011-01-01

    We have investigated rapid, label free detection of white spot syndrome virus (WSSV) using the first longitudinal extension resonance peak of five lead-magnesium niobate-lead titanate (PMN-PT) piezoelectric microcantilever sensors (PEMS) 1050-700 ?m long and 850-485 ?m wide constructed from 8 ?m thick PMN-PT freestanding films. The PMN-PT PEMS were encapsulated with a 3-mercaptopropltrimethoxysilane (MPS) insulation layer and further coated with anti-VP28 and anti-VP664 antibodies to target the WSSV virions and nucleocapsids, respectively. By inserting the antibody-coated PEMS in a flowing virion or nucleocapsid suspension, label-free detection of the virions and nucleocapsids were respectively achieved by monitoring the PEMS resonance frequency shift. We showed that positive label-free detection of both the virion and the nucleocapsid could be achieved at a concentration of 100 virions (nucleocapsids)/ml or 10 virions (nucleocapsids)/100?l, comparable to the detection sensitivity of polymerase chain reaction (PCR). However, in contrast to PCR, PEMS detection was label-free, in-situ and rapid (less than 30 min), potentially requiring minimal or no sample preparation. PMID:20863681

  13. [The use of therapeutic play in the intensive care of a preschool child with virus-associated hemophagocytic syndrome].

    PubMed

    Hsu, Chia-Hua; Feng, Jui-Ying

    2015-04-01

    Hospitalization is a stressful experience for children that increases their anxiety and fears, generates resistance and noncompliance, and, as a result, delays necessary treatments. Developing an age-appropriate intervention to reduce the hospitalization-related stress perceived by children is an important component of pediatric nursing. This case study used therapeutic play and drawing to care for a virus-associated hemophagocytic syndrome preschooler who stayed in our pediatric intensive care unit (PICU) between 11/13/2012 and 11/19/2012. Stressors faced by the patient included separation from primary caregiver, unfamiliarity with the medical environment and equipment, non-comprehension of the treatment and medication regimens, and loss of control. The patient displayed incorporative behaviors such as crying, screaming, refusing to be touched, and requesting parental accompaniment. Painting and picture books were used as developmentally appropriate interventions to understand the patient's feelings and to provide a means for him to project and release emotions. This strategy successfully assisted the child to overcome the perceived stress of hospitalization and to cooperate with healthcare providers on his treatment. PMID:25854953

  14. Proteomic alteration of Marc-145 cells and PAMs after infection by porcine reproductive and respiratory syndrome virus.

    PubMed

    Ding, Zhuang; Li, Zhi-jie; Zhang, Xiao-dong; Li, Ya-gang; Liu, Chang-jun; Zhang, Yan-Ping; Li, Yang

    2012-01-15

    Viral infections usually result in alterations in the host cell proteome, which determine the fate of infected cells and the progress of pathogenesis. To uncover cellular protein responses in porcine reproductive and respiratory syndrome virus (PRRSV), infected pulmonary alveolar macrophages (PAMs) and Marc-145 cells were subjected to proteomic analysis involving two-dimensional electrophoresis (2-DE) followed by MALDI-TOF-MS/MS identification. Altered expression of 44 protein spots in infected cells was identified in 2D gels, of which the 29 characterised by MALDI-TOF-MS/MS included 17 up-regulated and 12 down-regulated proteins. Some of these proteins were further confirmed at the mRNA level using real-time RT-PCR. Moreover, Western blot analysis confirmed the up-regulation of HSP27, vimentin and the down-regulation of galectin-1. Our study is the first attempt to analyze the cellular protein profile of PRRSV-infected Marc-145 cells using proteomics to provide valuable information about the effects of PRRSV-induced alterations on Marc-145 cell function. Further study of the affected proteins may facilitate our understanding of the mechanisms of PRRSV infection and pathogenesis. PMID:22137209

  15. Genetic diversity analysis of the ORF5 gene in porcine reproductive and respiratory syndrome virus samples from South China.

    PubMed

    Cao, Zong-Xi; Jiao, Pei-Rong; Huang, Yu-Mao; Qin, Hong-Yang; Kong, Liu-Wu; Pan, Quan-Hui; He, Yi-Min; Zhang, Gui-Hong

    2012-03-01

    To understand the genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) in South China, we collected 231 clinical samples from pigs with suspected PRRSV infection in Guangdong between 2007 and 2009. We found that 74 of 231 samples were positive by RT-PCR. The PCR products of the ORF5 gene of 35 isolates from different farms were sequenced and their DNA sequences were compared to 23 other PRRSV isolates in the GenBank. We found that the nucleotide similarity among all South China isolates ranged from 87.6% to 100%, and all belonged to the North American genotype. Most of them were classified into subgenotype I, but the rest mapped to subgenotypes III, V or VI. Those in subgenotypes I and III were found to be highly variable in the primary neutralising epitope (PNE) with a specific amino acid mutation (F39/L39?I39), and a few isolates in subgenotypes I and III isolates also had a mutation at L41 (L41?S41). PRRSV isolates in subgenotypes III, V and VI had less potential glycosylation sites than those in subgenotype I. Our data contribute to the understanding of molecular variation of PRRSV in South China. PMID:22366140

  16. Ebola Virus and Severe Acute Respiratory Syndrome Coronavirus Display Late Cell Entry Kinetics: Evidence that Transport to NPC1+ Endolysosomes Is a Rate-Defining Step

    PubMed Central

    Mingo, Rebecca M.; Simmons, James A.; Shoemaker, Charles J.; Nelson, Elizabeth A.; Schornberg, Kathryn L.; D'Souza, Ryan S.; Casanova, James E.

    2014-01-01

    ABSTRACT Ebola virus (EBOV) causes hemorrhagic fevers with high mortality rates. During cellular entry, the virus is internalized by macropinocytosis and trafficked through endosomes until fusion between the viral and an endosomal membrane is triggered, releasing the RNA genome into the cytoplasm. We found that while macropinocytotic uptake of filamentous EBOV viruslike particles (VLPs) expressing the EBOV glycoprotein (GP) occurs relatively quickly, VLPs only begin to enter the cytoplasm after a 30-min lag, considerably later than particles bearing the influenza hemagglutinin or GP from lymphocytic choriomeningitis virus, which enter through late endosomes (LE). For EBOV, the long lag is not due to the large size or unusual shape of EBOV filaments, the need to prime EBOV GP to the 19-kDa receptor-binding species, or a need for unusually low endosomal pH. In contrast, since we observed that EBOV entry occurs upon arrival in Niemann-Pick C1 (NPC1)-positive endolysosomes (LE/Lys), we propose that trafficking to LE/Lys is a key rate-defining step. Additional experiments revealed, unexpectedly, that severe acute respiratory syndrome (SARS) S-mediated entry also begins only after a 30-min lag. Furthermore, although SARS does not require NPC1 for entry, SARS entry also begins after colocalization with NPC1. Since the only endosomal requirement for SARS entry is cathepsin L activity, we tested and provide evidence that NPC1+ LE/Lys have higher cathepsin L activity than LE, with no detectable activity in earlier endosomes. Our findings suggest that both EBOV and SARS traffic deep into the endocytic pathway for entry and that they do so to access higher cathepsin activity. IMPORTANCE Ebola virus is a hemorrhagic fever virus that causes high fatality rates when it spreads from zoonotic vectors into the human population. Infection by severe acute respiratory syndrome coronavirus (SARS-CoV) causes severe respiratory distress in infected patients. A devastating outbreak of EBOV occurred in West Africa in 2014, and there was a significant outbreak of SARS in 2003. No effective vaccine or treatment has yet been approved for either virus. We present evidence that both viruses traffic late into the endocytic pathway, to NPC1+ LE/Lys, in order to enter host cells, and that they do so to access high levels of cathepsin activity, which both viruses use in their fusion-triggering mechanisms. This unexpected similarity suggests an unexplored vulnerability, trafficking to NPC1+ LE/Lys, as a therapeutic target for SARS and EBOV. PMID:25552710

  17. A transgenic Marc-145 cell line of piggyBac transposon-derived targeting shRNA interference against porcine reproductive and respiratory syndrome virus.

    PubMed

    Zhou, Fang; Liang, Shuang; Chen, An-hui; Singh, Chabungbam Orville; Bhaskar, Roy; Niu, Yan-shan; Miao, Yun-gen

    2012-06-01

    Porcine reproductive and respiratory syndrome (PRRS) is now considered to be one of the most important diseases in countries with intensive swine industries. The two major membrane-associated proteins of porcine reproductive and respiratory syndrome virus (PRRSV), GP5 and M (encoded by ORF5 and ORF6 genes, respectively), are associated as disulfide-linked heterodimers (GP5/M) in the virus particle. In this study, we designed 5 of the small hairpin RNAs (shRNAs) targeting the GP5 and M gene of PRRSV respectively, and investigated their inhibition to the production of PRRSV. The highest activity displayed in shRNAs of the ORF6e sequence (nts 261-279), which the inhibition rate reached was 99.09%. The result suggests that RNAi technology might serve as a potential molecular strategy for PRRSV therapy. Furthermore, the transgenic Marc-145 cell line of piggyBac transposon-derived targeting shRNA interference against PRRS virus was established. It presented stable inhibition to the replication and amplification of PRRS. The work implied that shRNAs targeting the GP5 and M gene of PRRSV may be used as potential RNA vaccines in vivo, and supplied the screening methods of transformed pig embryonic fibroblast which are prerequisite for the disease-resistant transgenic pigs to PRRS. PMID:22297554

  18. Evaluation of the presence of porcine reproductive and respiratory syndrome virus in pig meat and experimental transmission following oral exposure

    PubMed Central

    2004-01-01

    Abstract A study was performed to evaluate the presence of porcine reproductive and respiratory syndrome virus (PRRSV) in pig meat collected at slaughterhouses and its potential transmission to pigs via pig meat. A total of 1039 blood samples were collected from pigs upon their arrival at the abattoir. The following day, meat samples (n = 1027) were collected from the carcasses of these same pigs. Samples originated from 2 Canadian slaughterhouses, 1 situated in the province of Quebec and the other situated in the province of Manitoba. Serum samples were tested for antibodies to PRRSV and both serum and meat samples were also tested for PRRSV nucleic acid by polymerase chain reaction (PCR). Seropositivity to PRRSV for all serum samples was 74.3%. Furthermore 45 (4.3%) of the total serum samples and 19 (1.9%) of the 1027 meat samples were positive for PRRSV by PCR. Sequence analysis of open reading frame (ORF) 5 performed on 15 of the 19 PRRSV strains identified in pig meat indicated that 9 were field strains and 6 were vaccine-like (98% to 99.7% nucleotide homology with the Ingelvac RespPRRS/Repro vaccine). One of these 6 strains presented an intermediate 2-6-2 restriction fragment length polymorphism (RFLP) cut pattern and the others showed the characteristic 2-5-2 RFLP pattern of the vaccine strain. All strains sequenced were determined to be North American strains. In only 1 of the 19 PRRSV-positive meat samples could PRRSV be isolated. To test the potential infectivity of meat samples containing residual PRRSV, 11 of the PCR-positive meat samples (weighing 1.05 to 1.8 kg) were each used in feeding experiments of 2 PRRSV antibody-negative specific pathogen-free pigs of 9 wk of age. Samples were cut into several pieces and fed to each pair of pigs on 2 consecutive days. Each pig pair was housed in a separate cubicle and serum samples were collected at –7, 0, 7, 14, and 20 to 21 days post exposure. Seven pig pairs were found to be infected by PRRSV following ingestion of meat samples, including meat samples containing vaccine-like virus, as judged by the demonstration of PRRSV antibodies and/or PRRSV nucleic acid in the serum. In summary, the present study indicated that low residual quantities of PRRSV may be found in a small percentage of pig meat collected at slaugtherhouses. Furthermore, when this meat was fed raw to pigs in the experimental setting designed, pigs could be infected by PRRSV. PMID:15581220

  19. In vivo detection of porcine reproductive and respiratory syndrome virus RNA by in situ hybridization at different times postinfection.

    PubMed Central

    Sur, J H; Cooper, V L; Galeota, J A; Hesse, R A; Doster, A R; Osorio, F A

    1996-01-01

    We studied the distribution of porcine reproductive and respiratory syndrome virus (PRRSV) RNA in tissues by in situ hybridization at different times postinfection (p.i.). The probe used for in situ hybridization was prepared by reverse transcription of PRRSV RNA, followed by PCR amplification of the cDNA. The sequence amplified corresponded to 433 bp from PRRSV open reading frame 7, which is contained in the nucleocapsid protein gene and which is highly conserved in both European and American strains (H. Mardassi, L. Wilson, S. Mounir, and S. Dea, J. Clin. Microbiol. 32:2197-2203, 1994). An immunohistochemical technique was used to detect PRRSV antigen in tissue from virus-infected animals by using a monoclonal antibody specific for the PRRSV nucleocapsid protein (E.A. Nelson, J. Christopher-Hennings, T. Drew, G. Wensvoort, J.E. Collins, and D.A. Benfield, J. Clin. Microbiol. 31:3184-3189, 1993). The detection of PRRSV RNA was conducted in tissues of 6-week-old pigs that had been infected with one of three different field PRRSV isolates and collected at times ranging from 4 to 42 days p.i. Hybridization signals specific for PRRSV RNA were detected in lung, lymphoid tissues, alveolar macrophages (obtained by lavage at the time of necropsy), Peyer's patches, and kidney. The PRRSV-positive cells in these tissues appeared to be predominantly macrophages. In lung tissue we also obtained evidence suggesting the involvement of type II pneumocytes in the replication of PRRSV. During the acute period of infection there was a close correlation between the detection of RNA and the detection of nucleocapsid protein in individual cells. At later times p.i. (28 and 42 days p.i.), instead, more cells containing only PRRSV RNA than those containing PRRSV RNA and also expressing PRRSV nucleocapsid protein were detected. These results suggest that PRRSV RNA might persist in the tissues of infected animals for a longer time than PRRSV antigen expression. PMID:8862599

  20. Modeling the Transmission of Middle East Respirator Syndrome Corona Virus in the Republic of Korea

    PubMed Central

    Xia, Zhi-Qiang; Zhang, Juan; Xue, Ya-Kui; Sun, Gui-Quan; Jin, Zhen

    2015-01-01

    The 2015 epidemic of Middle East respiratory syndrome (MERS) in the Republic of Korea has been the largest outbreak outside Middle East. This epidemic had caused 185 laboratory-confirmed cases and 36 deaths in the Republic of Korea until September 2, 2015, which attracted public’s attention. Based on the detailed data of patients released by World Health Organization (WHO) and actual propagation of the epidemic, we construct two dynamical models to simulate the propagation processes from May 20 to June 8 and from June 9 to July 10, 2015, respectively and find that the basic reproduction number R0 reaches up to 4.422. The numerical analysis shows that the reasons of the outbreak spread quickly are lack of self-protection sense and targeted control measures. Through partial correction analysis, the parameters ?1 and ? have strong correlations with R0, i.e., the infectivity and proportion of the asymptomatic infected cases have much influence on the spread of disease. By sensitivity analysis, strengthening self-protection ability of susceptible and quickly isolating or monitoring close contacts are effective measures to control the disease. PMID:26690750

  1. Knowledge and attitude toward human immunodeficiency virus/acquired immuno deficiency syndrome among dental and medical undergraduate students

    PubMed Central

    Kumar, Vinod; Patil, Kavitha; Munoli, Karishma

    2015-01-01

    Background and Objectives: Human immunodeficiency virus (HIV) is a major public health challenge. Unjustified calls for the isolation of patients with HIV infection might further constrain the potential for expansion of clinical services to deal with a greater number of such patients. This infectious illness can evoke irrational emotions and fears in health care providers. Keeping this in view, a study was conducted to assess the knowledge and attitudes related to HIV/acquired immune deficiency syndrome (AIDS) among dental and medical students. Methodology: Descriptive cross-sectional survey of the entire dental and medical undergraduate students from two colleges was carried out using a pretested, self-administered questionnaire. Descriptive statistics such as percentage was used to present the data. Results: Ninety-eight percentage medical and dental undergraduate graduate students knew about HIV transmission in the hospital. Journals and internet were the leading source of information among both medical and dental undergraduates. The majority of respondents discussed HIV-related issues with their classmates. Surprisingly, 38% medical and 52% dental undergraduates think that HIV patient should be quarantined (isolation) to prevent the spread of infection. 68% medical and 60% dental undergraduates are willing to rendering dental/medical care to HIV-infected patients. Relatively large proportion (98%) of participants was willing to participate for HIV prevention program. Conclusion: The knowledge of medical and dental students is adequate, but the attitude needs improvement. Dental and medical students constitute a useful public health education resource. Comprehensive training, continuing education, and motivation will improve their knowledge and attitude, which enable them to provide better care to HIV patients. PMID:26538940

  2. Adeno-Associated Virus-Mediated Rescue of the Cognitive Defects in a Mouse Model for Angelman Syndrome

    PubMed Central

    Daily, Jennifer L.; Nash, Kevin; Jinwal, Umesh; Golde, Todd; Rogers, Justin; Peters, Melinda M.; Burdine, Rebecca D.; Dickey, Chad; Banko, Jessica L.; Weeber, Edwin J.

    2011-01-01

    Angelman syndrome (AS), a genetic disorder occurring in approximately one in every 15,000 births, is characterized by severe mental retardation, seizures, difficulty speaking and ataxia. The gene responsible for AS was discovered to be UBE3A and encodes for E6-AP, an ubiquitin ligase. A unique feature of this gene is that it undergoes maternal imprinting in a neuron-specific manner. In the majority of AS cases, there is a mutation or deletion in the maternally inherited UBE3A gene, although other cases are the result of uniparental disomy or mismethylation of the maternal gene. While most human disorders characterized by severe mental retardation involve abnormalities in brain structure, no gross anatomical changes are associated with AS. However, we have determined that abnormal calcium/calmodulin-dependent protein kinase II (CaMKII) regulation is seen in the maternal UBE3A deletion AS mouse model and is responsible for the major phenotypes. Specifically, there is an increased ?CaMKII phosphorylation at the autophosphorylation sites Thr286 and Thr305/306, resulting in an overall decrease in CaMKII activity. CaMKII is not produced until after birth, indicating that the deficits associated with AS are not the result of developmental abnormalities. The present studies are focused on exploring the potential to rescue the learning and memory deficits in the adult AS mouse model through the use of an adeno-associated virus (AAV) vector to increase neuronal UBE3A expression. These studies show that increasing the levels of E6-AP in the brain using an exogenous vector can improve the cognitive deficits associated with AS. Specifically, the associative learning deficit was ameliorated in the treated AS mice compared to the control AS mice, indicating that therapeutic intervention may be possible in older AS patients. PMID:22174738

  3. Human immunodeficiency virus disease in California. Effects of the 1993 expanded case definition of the acquired immunodeficiency syndrome.

    PubMed Central

    Singleton, J A; Tabnak, F; Kuan, J; Rutherford, G W

    1996-01-01

    On January 1, 1993, the case definition of the acquired immunodeficiency syndrome (AIDS) in adults and adolescents used for monitoring the AIDS epidemic in California was expanded to include persons infected with the human immunodeficiency virus (HIV) with CD4 T-lymphocyte counts of less than 200 x 10(6) per liter (< 200 per mm3), pulmonary tuberculosis, recurrent pneumonia, or invasive cervical cancer. To assess the implications of this revision on AIDS case reporting in California, we compared cases reported through the end of 1994 based on 1 or more of the 4 new AIDS-defining conditions added in 1993 to cases reported based on pre-1993 AIDS-defining opportunistic infections and cancers. The 4 new conditions included in the 1993 expanded AIDS case definition accounted for a 23% increase in cumulative AIDS cases reported in California by the end of 1993, a 170% increase in the number of cases reported during 1993, and an 88% increase in the number of patients with AIDS living at the end of 1993. The number of cases reported in 1993 (19,629) was 124% more than that reported in 1992 (8,780) and 69% more than that reported in 1994 (11,587). The proportion of cases among women, injection-drug users, and African Americans also increased as a result of this change in the case definition. The expansion of the case definition may have resulted in a peak or plateau in the AIDS incidence in California because of reporting earlier in the HIV disease progression. The expanded case definition has enhanced the usefulness of AIDS surveillance data for targeting secondary prevention efforts, but more behavioral and HIV serosurveys are still needed to adequately target primary HIV prevention efforts. Images Figure 1. PMID:8775725

  4. Immunogenicity of recombinant GP5 protein of porcine reproductive and respiratory syndrome virus expressed in tobacco plant.

    PubMed

    Chia, Min-Yuan; Hsiao, Shih-Hsuan; Chan, Hui-Ting; Do, Yi-Yin; Huang, Pung-Ling; Chang, Hui-Wen; Tsai, Yi-Chieh; Lin, Chun-Ming; Pang, Victor Fei; Jeng, Chian-Ren

    2010-06-15

    The aim of the study was to evaluate the immunogenicity of the ORF5-encoded major envelop glycoprotein 5 (GP5) of porcine reproductive and respiratory syndrome virus (PRRSV) expressed in tobacco plant as a potential pig oral vaccine in protection against PRRSV infection. Six-week-old PRRSV-free pigs were fed four times orally with 50g of chopped fresh GP5 transgenic tobacco leaves (GP5-T) (GP5 reaching 0.011% of total soluble protein) or wild-type tobacco leaves (W-T) each on days 0, 14, 28, and 42. Samples of serum, saliva, and peripheral blood mononuclear cells (PBMCs) were collected on days -1, 6, 13, 20, 27, 34, 41, and 48 after the initial oral vaccination. A similar vaccination-dependent gradual increase in the responses of serum and saliva anti-PRRSV total IgG and IgA, respectively, and in the levels of PRRSV-specific blastogenic response of PBMCs was seen in GP5-T-treated pigs; all statistically significant elevations occurred after the 2nd vaccination and were revealed after 20 days post-initial oral vaccination (DPIOV). Pigs fed on GP5-T also developed serum neutralizing antibodies to PRRSV at a titer of 1:4-1:8 after the 4th vaccination by 48 DPIOV. No detectable anti-PRRSV antibody responses and PRRSV-specific blastogenic response were seen in W-T-treated pigs. The present study has demonstrated that pigs fed on GP5-T could develop specific mucosal as well as systemic humoral and cellular immune responses against PRRSV. The results also support that transgenic plant as GP5-T can be an effective system for oral delivery of recombinant subunit vaccines in pigs. PMID:20053461

  5. Porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 infections in wild boar (Sus scrofa) in southwestern Germany.

    PubMed

    Hammer, Ralf; Ritzmann, Mathias; Palzer, Andreas; Lang, Christiane; Hammer, Birgit; Pesch, Stefan; Ladinig, Andrea

    2012-01-01

    Samples were collected from 203 wild boars (Sus scrofa) hunted in Baden-Wurtemburg, Germany from November-January 2008 and 2009. Samples from the lung and tonsil were analyzed by quantitative polymerase chain reaction (qPCR) for porcine reproductive and respiratory syndrome virus (PRRSV) type 1 (European type) and type 2 (American type). A qPCR to detect porcine circovirus type 2 (PCV2)-specific genome was performed on tissue homogenates including lung, tonsils, and inguinal lymph nodes. Serum samples were tested for antibodies against PRRSV and PCV2 by enzyme-linked immunosorbent assay (ELISA). No PRRSV was detected in any of the 203 samples and one sample had detectable antibodies against PRRSV. We detected PCV2 in organ materials from 103 wild boars with a prevalence of 50.7%. The number of wild boars positive for PCV2 by PCR varied according to the population density of wild boars among woodlands. More positive samples were detected in woodlands with a high density of wild boars. We found no correlation between the number of PCV2-positive wild boars and the density of domestic pigs in the surrounding area. The number of wild boars positive for antibodies against PCV2 by the INGEZIM Circovirus IgG/IgM test kit was low (53 sera positive for IgG- and three sera positive for IgM-antibodies) in comparison to the higher positive results from the INGEZIM CIRCO IgG test kit (102 positive and 12 inconclusive results). PMID:22247377

  6. Effects on boar semen quality after infection with porcine reproductive and respiratory syndrome virus: a case report.

    PubMed

    Schulze, Martin; Revilla-Fernández, Sandra; Schmoll, Friedrich; Grossfeld, Rudolf; Griessler, Alfred

    2013-01-01

    The effect of porcine reproductive and respiratory syndrome virus (PRRSV) on semen quality was examined in a group of 11 spontaneously infected boars in a commercial boar stud. Semen samples were collected 4 weeks prior to 4 weeks post-infection (wpi). Infection with PRRSV of the European genotype subtype 1 (EU-1) was verified by specific quantitative real-time polymerase chain reaction (RT-PCR) in 36% of the serum samples. All boars seroconverted before 4 wpi and remained in normal condition throughout the study. Comparison of the percentage of morphologically intact spermatozoa revealed an increase of acrosome-defective spermatozoa (P = 0.012) between -4 and 4 wpi. Significant deleterious effects on semen quality were detected for membrane integrity when semen had been stored for 2 days after sampling. Analysis of sperm subpopulations in a thermoresistance test on day 7 after sampling revealed alterations in the percentage of circular, progressively motile spermatozoa (P = 0.013), in the percentage of non-linear, progressively motile spermatozoa (P = 0.01), and on the amplitude of lateral sperm head displacement (P = 0.047). There was no difference in the incidence of mitochondrially active spermatozoa (P = 0.075). Investigation of routine production data between pre- and post-infection status showed no differences on ejaculate volume (P = 0.417), sperm concentration (P = 0.788), and percentage of motile spermatozoa (P = 0.321). This case report provides insights into a potential control strategy for PRRSV outbreaks in boar studs. PMID:23442207

  7. High infection rate of bank voles (Myodes glareolus) with Puumala virus is associated with a winter outbreak of haemorrhagic fever with renal syndrome in Croatia.

    PubMed

    Tadin, A; Bjedov, L; Margaletic, J; Zibrat, B; Krajinovic, L Cvetko; Svoboda, P; Kurolt, I C; Majetic, Z Stritof; Turk, N; Rode, O Dakovic; Civljak, R; Kuzman, I; Markotic, A

    2014-09-01

    An outbreak of haemorrhagic fever with renal syndrome (HFRS) started on Medvednica mountain near Zagreb in January 2012. In order to detect the aetiological agent of the disease in small rodents and to make the link with the human outbreak, rodents were trapped at four different altitudes. Using nested RT-PCR, Puumala virus (PUUV) RNA was detected in 41/53 (77·4%) bank voles (Myodes glareolus) and Dobrava virus (DOBV) RNA was found in 6/61 (9·8%) yellow-necked mice (Apodemus flavicollis). Sequence analysis of a 341-nucleotide region of the PUUV S segment, obtained from all infected bank voles and five HFRS patients, showed 98·8-100% sequence similarity, indicating that the patients were probably exposed to PUUV on Medvednica mountain. A very large bank-vole population combined with an extremely high infection rate of PUUV was responsible for this unusual winter outbreak of HFRS in Croatia. PMID:24800636

  8. Development and evaluation of a novel reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of type II porcine reproductive and respiratory syndrome virus.

    PubMed

    Gao, Ming; Cui, Jin; Ren, Yudong; Suo, Siqingaowa; Li, Guangxing; Sun, Xuejiao; Su, Dingding; Opriessnig, Tanja; Ren, Xiaofeng

    2012-10-01

    The objective of this study was to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of type II porcine reproductive and respiratory syndrome virus (PRRSV). Based on sequence alignment, four primers were designed amplifying the M gene of type II PRRSV and were subsequently utilized in an RT-LAMP assay. The RT-LAMP product had a ladder-like pattern of bands and the optimal reaction condition for this assay was determined to be 40 min at 63°C. Comparative analysis indicated that the RT-LAMP method was more sensitive than a conventional RT-PCR assay and comparable to a real-time PCR assay. In addition, the RT-LAMP assay was capable of detecting type II PRRSV in field samples and differentiating type II PRRSV from seven other porcine viruses which are all associated frequently with similar clinical symptoms. PMID:22659065

  9. Nsp9 and Nsp10 Contribute to the Fatal Virulence of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Emerging in China

    PubMed Central

    Zhang, Jialong; Ge, Xinna; Zhou, Rong; Zheng, Huaguo; Geng, Gang; Guo, Xin; Yang, Hanchun

    2014-01-01

    Atypical porcine reproductive and respiratory syndrome (PRRS), which is caused by the Chinese highly pathogenic PRRS virus (HP-PRRSV), has resulted in large economic loss to the swine industry since its outbreak in 2006. However, to date, the region(s) within the viral genome that are related to the fatal virulence of HP-PRRSV remain unknown. In the present study, we generated a series of full-length infectious cDNA clones with swapped coding regions between the highly pathogenic RvJXwn and low pathogenic RvHB-1/3.9. Next, the in vitro and in vivo replication and pathogenicity for piglets of the rescued chimeric viruses were systematically analyzed and compared with their backbone viruses. First, we swapped the regions including the 5?UTR+ORF1a, ORF1b, and structural proteins (SPs)-coding region between the two viruses and demonstrated that the nonstructural protein-coding region, ORF1b, is directly related to the fatal virulence and increased replication efficiency of HP-PRRSV both in vitro and in vivo. Furthermore, we substituted the nonstructural protein (Nsp) 9-, Nsp10-, Nsp11- and Nsp12-coding regions separately; or Nsp9- and Nsp10-coding regions together; or Nsp9-, Nsp10- and Nsp11-coding regions simultaneously between the two viruses. Our results indicated that the HP-PRRSV Nsp9- and Nsp10-coding regions together are closely related to the replication efficiency in vitro and in vivo and are related to the increased pathogenicity and fatal virulence for piglets. Our findings suggest that Nsp9 and Nsp10 together contribute to the fatal virulence of HP-PRRSV emerging in China, helping to elucidate the pathogenesis of this virus. PMID:24992286

  10. Prevalence of porcine reproductive and respiratory syndrome virus detection in aborted fetuses, mummified fetuses and stillborn piglets using quantitative polymerase chain reaction

    PubMed Central

    OLANRATMANEE, Em-on; WONGYANIN, Piya; THANAWONGNUWECH, Roongroje; TUMMARUK, Padet

    2015-01-01

    The objective of the present study was to investigate the prevalence of porcine reproductive and respiratory syndrome (PRRS) virus detection in aborted fetuses (n=32), mummified fetuses (n=30) and stillborn piglets (n=27) from 10 swine herds in Thailand using quantitative polymerase chain reaction (qPCR). Pooled organs and umbilical cord from each fetus/piglet were homogenized and subjected to RNA extraction and cDNA synthesis. The qPCR was carried out on the ORF7 of the PRRS viral genome using fluorogenic probes for amplified product detection. The results revealed that 67.4% (60/89) of the specimens contained PRRS virus. The virus was found in 65.6% (21/32) of aborted fetuses, 63.3% (19/30) of mummified fetuses and 74.1% (20/27) of stillborn piglets (P=0.664). Genotype 1, genotype 2 and mixed genotypes of PRRS virus were detected in 19.1% (17/89), 25.8% (23/89) and 22.5% (20/89) of the specimens, respectively (P=0.316). PRRS virus antigen was retrieved from both non-PRRS-vaccinated herds (68.2%, 45/66) and PRRS-vaccinated herds (65.2%, 15/23) (P=0.794). These findings indicated that these specimens are important sources of the PRRS viral load and the viral shedding within the herd. Thus, intensive care on the routine management of dead fetuses and stillborn piglets in PRRS virus-positive herds should be emphasized. PMID:25866409

  11. The impact of porcine reproductive and respiratory syndrome virus genetic heterogeneity on molecular assay performances.

    PubMed

    Drigo, Michele; Franzo, Giovanni; Gigli, Alessandra; Martini, Marco; Mondin, Alessandra; Gracieux, Patrice; Ceglie, Letizia

    2014-06-01

    The remarkable economic losses due to porcine reproductive and respiratory syndrome (PRRS) have stated the control and eradication of this disease is one of the main issues of swine modern farming. The limited cross-protection of vaccine-induced immunity compelled the adoption of strict biosecurity measures that must be associated with the prompt diagnosis of infection. In our study four RT-PCR methods, a RT-PCR, a SYBR Green I and two hydrolysis probes, were compared to evaluate their respective benefits and disadvantages. One hundred and seventy samples originating from 50 farms located in northern Italy were tested with all assays and performances were evaluated using a Bayesian approach to deal with the absence of a Gold Standard. Sequencing the complete of ORF7, the segment targeted by all methods, allowed a gain of insight into the genetic variability of Italian strains and to investigate the role of mismatches on assay sensitivity. Our study evidenced that methods based only on primers-genome interaction better tolerate PRRSV genetic variability, demonstrating a greater sensitivity (Se): SYBR Green I (Se=98.4%) and RT-PCR (Se=99%) outperform both in-house (Se=71.4%) and commercial (Se=91.7%) probe-based methods. On the other hand, probe-based assays allowed an easier genotyping of PRRSV strains and implementation of the internal control system (IC). Phylogenetic analysis allowed demonstration of a presence of two clades circulating continuously in northern Italy since 1996, when their probable ancestors were collected. PMID:24642238

  12. Concise and broadly applicable method for determining the genomic sequences of North-American-type porcine reproductive and respiratory syndrome viruses in various clusters.

    PubMed

    Morozumi, Takeya; Iseki, Hiroshi; Toki, Daisuke; Takagi, Michihiro; Tsunemitsu, Hiroshi; Uenishi, Hirohide

    2014-09-01

    We developed a concise and broadly applicable method for accurate genomic sequencing of North American genotype (NA-type) porcine reproductive and respiratory syndrome viruses (PRRSVs) that overcomes high genetic variability of the viruses. The method, designated "combination of consensus oligonucleotide reverse transcription and multiple displacement amplification" (CORT-MDA), involves reverse-transcription of viral RNA followed by shotgun sequencing after amplification using only 11 degenerate oligonucleotide primers; these primers were designed against consensus regions within the open reading frames of the 124 NA-type PRRSV strains with reported full-length genomic sequences. Sequencing of the 192 shotgun clones generated per virus showed 80% to 94% coverage on the reported PRRSV genomic sequence, such that only 2 or 3 unread regions had to be resequenced after PCR amplification using custom primers. Direct sequencing of RT-PCR products confirmed absolute consistency between sequences determined by the CORT-MDA method and those from RT-PCR. These results suggest that our method is applicable to diverse NA-type viruses. PMID:24920486

  13. 22 CFR 72.14 - Nominal possession; property not normally taken into physical possession.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...Nominal possession; property not normally taken into physical possession. ...Nominal possession; property not normally taken into physical possession. ...expected to take physical possession of items of personal property such as:...

  14. 22 CFR 72.14 - Nominal possession; property not normally taken into physical possession.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...Nominal possession; property not normally taken into physical possession. ...Nominal possession; property not normally taken into physical possession. ...expected to take physical possession of items of personal property such as:...

  15. 22 CFR 72.14 - Nominal possession; property not normally taken into physical possession.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...Nominal possession; property not normally taken into physical possession. ...Nominal possession; property not normally taken into physical possession. ...expected to take physical possession of items of personal property such as:...

  16. 22 CFR 72.14 - Nominal possession; property not normally taken into physical possession.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...Nominal possession; property not normally taken into physical possession. ...Nominal possession; property not normally taken into physical possession. ...expected to take physical possession of items of personal property such as:...

  17. 22 CFR 72.14 - Nominal possession; property not normally taken into physical possession.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...Nominal possession; property not normally taken into physical possession. ...Nominal possession; property not normally taken into physical possession. ...expected to take physical possession of items of personal property such as:...

  18. Assessment of the Safety and Efficacy of an Attenuated Live Vaccine Based on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus

    PubMed Central

    Yu, Xiuling; Zhou, Zhi; Cao, Zhen; Wu, Jiajun; Zhang, Zhongqiu; Xu, Baiwan; Wang, Chuanbin; Hu, Dongmei; Deng, Xiaoyu; Han, Wei; Gu, Xiaoxue; Zhang, Shuo; Li, Xiaoxia; Wang, Baoyue

    2015-01-01

    The safety and efficacy of the JXA1-R vaccine, an attenuated strain of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV), were examined using an intramuscular challenge model in piglets. The JXA1-R vaccine was obtained by passing HP-PRRSV JXA1 through Marc-145 cells (82nd passage). Genomic sequence comparisons showed that strain JXA1-R and its parental strain, JXA1, differ by 47 amino acids, and most of these differences are scattered throughout the PRRSV genome. Four-week-old PRRSV-free piglets were inoculated intramuscularly with JXA1-R vaccine (103.0, 104.0, 105.0, 106.0, and 107.0 50% tissue culture infective doses [TCID50]/ml for groups 1 to 5, respectively) and then challenged intramuscularly with the 5th passage virus of JXA1 virus (JXA1-F5, 3 ml × 104.5 TCID50/ml) 28 days after inoculation. The humoral immune response, swine growth, clinical signs, and differential organ lesions were monitored. The results showed that all vaccinated piglets had a perceptible humoral immune response to vaccination after day 7, which then promptly increased, almost reaching the maximum sample/positive (S/P) ratio value at 28 days postimmunization. Viremia detection indicated that the viral replication levels of the challenge virus in the immunized groups (immunization doses ?104.0/ml) were significantly lower than that of the virus-challenged unvaccinated control group. Piglets in groups 2 to 5 were effectively protected against lethal HP-PRRSV infection and did not show any obvious changes in body temperature or clinical signs of disease at any point during the experiment. However, two of five piglets in group 1 showed mild pathological lesions and transitory high fever. These results suggest that JXA1-R (TCID50/ml ?104.0) is sufficiently attenuated and can provide effective protection against the lethal wild-type HP-PRRSV. PMID:25739919

  19. The 30-Amino-Acid Deletion in the Nsp2 of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Emerging in China Is Not Related to Its Virulence?

    PubMed Central

    Zhou, Lei; Zhang, Jialong; Zeng, Jingwen; Yin, Shuoyan; Li, Yanhua; Zheng, Linying; Guo, Xin; Ge, Xinna; Yang, Hanchun

    2009-01-01

    During the past 2 years, an atypical clinical outbreak, caused by a highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) with a unique 30-amino-acid deletion in its Nsp2-coding region, was pandemic in China. In this study, we generated four full-length infectious cDNA clones: a clone of the highly virulent PRRSV strain JXwn06 (pWSK-JXwn), a clone of the low-virulence PRRSV strain HB-1/3.9 (pWSK-HB-1/3.9), a chimeric clone in which the Nsp2 region containing the 30-amino-acid deletion was replaced by the corresponding region of the low-virulence PRRSV strain HB-1/3.9 (pWSK-JXwn-HB1nsp2), and a mutated HB-1/3.9 clone with the same deletion in Nsp2 as JXwn06 (pWSK-HB1-ND30). We also investigated the pathogenicities of the rescued viruses (designated RvJXwn, RvJXwn-HB1nsp2, RvHB-1/3.9, and RvHB1-ND30, respectively) in specific-pathogen-free piglets in order to determine the role of the 30-amino-acid deletion in the virulence of the highly pathogenic PRRSV. All the rescued viruses could replicate stably in MARC-145 cells. Our findings indicated that RvJXwn-HB1nsp2 retained high virulence for piglets, like RvJXwn and the parental virus JXwn06, although the survival time of piglets infected with RvJXwn-HB1nsp2 was obviously prolonged. RvHB1-ND30 exhibited low virulence for piglets, like RvHB-1/3.9 and the parental virus HB-1/3.9. Therefore, we conclude that the 30-amino-acid deletion is not related to the virulence of the highly pathogenic PRRSV emerging in China. PMID:19244318

  20. Testing for Human Immunodeficiency Virus

    MedlinePLUS

    ... incisions made in the mother’s abdomen and uterus. Human Immunodeficiency Virus (HIV): A virus that attacks certain cells of the body’s immune system and causes acquired immunodeficiency syndrome (AIDS). Immune System: ...

  1. Yeast Surface Display of Two Proteins Previously Shown to Be Protective Against White Spot Syndrome Virus (WSSV) in Shrimp

    PubMed Central

    Ananphongmanee, Vorawit; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Boonchird, Chuenchit

    2015-01-01

    Cell surface display using the yeasts Saccharomyces cerevisiae and Pichia pastoris has been extensively developed for application in bioindustrial processes. Due to the rigid structure of their cell walls, a number of proteins have been successfully displayed on their cell surfaces. It was previously reported that the viral binding protein Rab7 from the giant tiger shrimp Penaeus monodon (PmRab7) and its binding partner envelope protein VP28 of white spot syndrome virus (WSSV) could independently protect shrimp against WSSV infection. Thus, we aimed to display these two proteins independently on the cell surfaces of 2 yeast clones with the ultimate goal of using a mixture of the two clones as an orally deliverable, antiviral agent to protect shrimp against WSSV infection. PmRab7 and VP28 were modified by N-terminal tagging to the C-terminal half of S. cerevisiae ?-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1) promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7) and partial VP28 (pVP28) were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA) using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against WSSV by oral administration. PMID:26083446

  2. Evaluation of the Efficacy of an Attenuated Live Vaccine against Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus in Young Pigs

    PubMed Central

    Leng, Xue; Li, Zhenguang; Xia, Mingqi; He, Yanliang

    2012-01-01

    Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is characterized by high fever and high mortality in pigs of all ages and has severely affected the pork industry of China in the last few years. An attenuated HP-PRRSV strain, TJM, was obtained by passaging HP-PRRSV strain TJ on MARC-145 cells for 92 passages. Porcine reproductive and respiratory syndrome virus (PRRSV)- and antibody-free pigs were inoculated intramuscularly with TJM (105.0 50% tissue culture infective doses [TCID50]) and challenged at 28, 60, 120, and 180 days postimmunization (dpi). The results showed that 5/5, 5/5, 5/5, and 4/5 immunized pigs were protected from the lethal challenge and did not develop fever and clinical diseases at each challenge, respectively. Compared to control pigs, vaccinated pigs showed much milder pathological lesions and gained significantly more weight (P < 0.01). Sequence analysis of different passages of strain TJ showed that the attenuation resulted in a deletion of a continuous 120 amino acids (aa), in addition to the discontinuous 30-aa deletion in the nsp2 region. The analysis also demonstrated that the 120-aa deletion was genetically stable in vivo. These results suggested that HP-PRRSV TJM was efficacious against a lethal challenge with a virulent HP-PRRSV strain, and effective protection could last at least 4 months. Therefore, strain TJM is a good candidate for an efficacious modified live virus vaccine as well as a useful molecular marker vaccine against HP-PRRSV. PMID:22695163

  3. Comparison of viremia of type II porcine reproductive and respiratory syndrome virus in naturally infected pigs by zip nucleic acid probe-based real-time PCR

    PubMed Central

    2013-01-01

    Background Porcine reproductive and respiratory syndrome virus (PRRSV) is a RNA virus with high genetic variation. This virus causes significant economic losses in most pig-producing countries. The clinical presentation of PRRSV ranges from asymptomatic to devastating. In this study, we developed a sensitive and specific zip nucleic acid probe-based real-time PCR assay to evaluate the viremia of natural PRRSV-infected pigs in Taiwan. Serum samples were collected from 577 pigs aged 5–12 weeks. These include 444 clinically healthy pigs and 133 symptomatic pigs were confirmed to have porcine respiratory disease complex (PRDC). Results Viremia was quantified in 79 of the 444 (17.8%) clinically healthy pigs and in 112 of the 133 (84.2%) PRDC cases. Viremias were significantly more common in pigs with PRDC compared with the clinically healthy pigs (P <0.0001). These results suggest that a high viral load is a major feature of PRRSV-affected pigs. Conclusions ZNA probe-based real-time PCR can be a useful tool to diagnose symptomatic and asymptomatic PRRSV-infected pigs. The presence of this marker in a sample of animals with high PRRSV loads (>104.2 PRRSV genomes/?l of serum) seems to indicate that it correlates with the presence of PRDC in pigs. PMID:24028493

  4. Comparative Analysis of Immune Responses in Pigs to High and Low Pathogenic Porcine Reproductive and Respiratory Syndrome Viruses Isolated in China.

    PubMed

    Han, Z; Liu, Y; Wang, G; He, Y; Hu, S; Li, Y; Shi, W; Wu, J; Wang, S; Liu, H; Cai, X

    2015-10-01

    The CH-1a and HuN4 strains of porcine reproductive and respiratory syndrome virus (PRRSV) show different pathogenicities in pigs. To understand host immune responses against these viruses, we investigated the dynamic changes in cytokine levels produced in peripheral blood of piglets infected with the highly pathogenic PRRSV HuN4 strain or the CH-1a strain. Clinical signs, virus loads and serum cytokine levels [interferon(IFN)-?, Interleukin (IL)-1, TNF-?, IL-6, IL-12, IFN-?, IL-10 and TGF-?] were tested. The results showed that while piglets developed effective cellular immune responses against CH-1a infection, those infected with HuN4 displayed ineffective cellular immunity, organ lesions and persistent elevated levels of immunoregulatory cytokines (IL-10 and TGF-?), which delayed the development of PRRSV-specific immune responses. These results demonstrated that HuN4 infection induced higher cytokine levels than that of CH-1a infection induced. The changes in inflammatory cytokines intensified the inflammatory reaction and damaged the tissues and organs. PMID:24308664

  5. Phylogeny-Directed Search for Murine Leukemia Virus-Like Retroviruses in Vertebrate Genomes and in Patients Suffering from Myalgic Encephalomyelitis/Chronic Fatigue Syndrome and Prostate Cancer

    PubMed Central

    Blomberg, Jonas; Sheikholvaezin, Ali; Elfaitouri, Amal; Blomberg, Fredrik; Sjösten, Anna; Mattson Ulfstedt, Johan; Pipkorn, Rüdiger; Källander, Clas; Öhrmalm, Christina; Sperber, Göran

    2011-01-01

    Gammaretrovirus-like sequences occur in most vertebrate genomes. Murine Leukemia Virus (MLV) like retroviruses (MLLVs) are a subset, which may be pathogenic and spread cross-species. Retroviruses highly similar to MLLVs (xenotropic murine retrovirus related virus (XMRV) and Human Mouse retrovirus-like RetroViruses (HMRVs)) reported from patients suffering from prostate cancer (PC) and myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) raise the possibility that also humans have been infected. Structurally intact, potentially infectious MLLVs occur in the genomes of some mammals, especially mouse. Mouse MLLVs contain three major groups. One, MERV G3, contained MLVs and XMRV/HMRV. Its presence in mouse DNA, and the abundance of xenotropic MLVs in biologicals, is a source of false positivity. Theoretically, XMRV/HMRV could be one of several MLLV transspecies infections. MLLV pathobiology and diversity indicate optimal strategies for investigating XMRV/HMRV in humans and raise ethical concerns. The alternatives that XMRV/HMRV may give a hard-to-detect “stealth” infection, or that XMRV/HMRV never reached humans, have to be considered. PMID:22315600

  6. Evaluation of Flinders Technology Associates cards for collection and transport of samples for detection of Porcine reproductive and respiratory syndrome virus by reverse transcription polymerase chain reaction.

    PubMed

    Linhares, Daniel C L; Rovira, Albert; Torremorell, Montserrat

    2012-03-01

    Blood, tissue and oral fluid samples collected from experimentally infected animals and field cases were used to evaluate the safety, diagnostic sensitivity and specificity of Flinders Technology Associates (FTA) cards for Porcine reproductive and respiratory syndrome virus (PRRSV) reverse transcription polymerase chain reaction (RT-PCR) diagnostics. The analytical sensitivity of PRRSV RT-PCR from serum and oral fluids in FTA cards was reduced, although the virus could still be detected at concentrations of 10(1) and 10(3) TCID/ml, respectively. The sensitivity and specificity of PRRSV RT-PCR detection from serum, blood, and tissue samples in cards collected from experimentally infected animals were 100%. Sensitivity for oral fluids was 45% (95% CI: 19.97-73.01) compared to fresh. For field samples, sensitivity was 89% (95% CI: 77.35-95.63) and 100% (95% CI: 80.00-100) for serum and lung samples, respectively. The sensitivity was the same for samples stored in cards at room temperature or at 4ºC, and tested overnight or after 14 days. Cards inoculated with PRRSV-positive samples did not yield replicating virus after cell culture. In conclusion, FTA cards proved to be a safe, simple, and sensitive alternative method to transport serum, blood, and tissue samples for PRRSV RT-PCR diagnostics; however, a significant decrease in RT-PCR sensitivity should be expected from oral fluid samples. PMID:22362527

  7. Development of a swine specific 9-plex Luminex cytokine assay and assessment of immunity after porcine reproductive and respiratory syndrome virus (PRRSV) vaccination: Elevated serum IL-12 levels are not predictive of protect

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A Luminex multiplex swine cytokine assay was developed to measure 9 cytokines simultaneously in pig serum and tested in a porcine reproductive and respiratory syndrome virus (PRRSV) vaccine/challenge study. This assay detects innate (IL-1ß, IL-6, IL-8, IFNa, TNFa); regulatory (IL-10), Th1 (IL-12, I...

  8. Porcine Reproductive and Respiratory Syndrome Virus: An update on an emerging and re-emerging viral disease of swine.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recognized in the late 1980’s in North America and Europe the syndrome that caused reproductive and respiratory problems in swine was initially called “Mystery Swine Disease” and is now termed “Porcine Reproductive and Respiratory Syndrome (PRRS)”. In the early 1990’s an arterivirus, referred to as ...

  9. An innovative approach to induce cross-protective immunity against porcine reproductive and respiratory syndrome virus in the lungs of pigs through adjuvanted nanotechnology-based vaccination

    PubMed Central

    Binjawadagi, Basavaraj; Dwivedi, Varun; Manickam, Cordelia; Ouyang, Kang; Torrelles, Jordi B; Renukaradhya, Gourapura J

    2014-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is an economically devastating respiratory disease of pigs. The disease is caused by the PRRS virus (PRRSV), an Arterivirus which is a highly mutating RNA virus. Widely used modified live PRRSV vaccines have failed to prevent PRRS outbreaks and reinfections; moreover, safety of the live virus vaccines is questionable. Though poorly immunogenic, inactivated PRRSV vaccine is safe. The PRRSV infects primarily the lung macrophages. Therefore, we attempted to strengthen the immunogenicity of inactivated/killed PRRSV vaccine antigens (KAg), especially in the pig respiratory system, through: 1) entrapping the KAg in biodegradable poly(lactic-co-glycolic acid) nanoparticles (NP-KAg); 2) coupling the NP-KAg with a potent mucosal adjuvant, whole cell lysate of Mycobacterium tuberculosis (M. tb WCL); and 3) delivering the vaccine formulation twice intranasally to growing pigs. We have previously shown that a single dose of NP-KAg partially cleared the challenged heterologous PRRSV. Recently, we reported that NP-KAg coupled with unentrapped M. tb WCL significantly cleared the viremia of challenged heterologous PRRSV. Since PRRSV is primarily a lung disease, our goal in this study was to investigate lung viral load and various immune correlates of protection at the lung mucosal surfaces and its parenchyma in vaccinated heterologous PRRSV-challenged pigs. Our results indicated that out of five different vaccine-adjuvant formulations, the combination of NP-KAg and unentrapped M. tb WCL significantly cleared detectable replicating infective PRRSV with a tenfold reduction in viral RNA load in the lungs, associated with substantially reduced gross and microscopic lung pathology. Immunologically, strong humoral (enhanced virus neutralization titers by high avidity antibodies) and cell-mediated immune responses (augmented population of interferon-? secreting CD4+ and CD8+ lymphocytes and reduced secretion of immunosuppressive cytokines) in the lungs were observed. In conclusion, combination of NP-KAg and soluble M. tb WCL elicits broadly cross-protective anti-PRRSV immunity in the pig respiratory system. PMID:24711701

  10. Development of a network based model to simulate the between-farm transmission of the porcine reproductive and respiratory syndrome virus.

    PubMed

    Thakur, Krishna K; Sanchez, Javier; Hurnik, Daniel; Poljak, Zvonimir; Opps, Sheldon; Revie, Crawford W

    2015-11-18

    Contact structure within a population can significantly affect the outcomes of infectious disease spread models. The objective of this study was to develop a network based simulation model for the between-farm spread of porcine reproductive and respiratory syndrome virus to assess the impact of contact structure on between-farm transmission of PRRS virus. For these farm level models, a hypothetical population of 500 swine farms following a multistage production system was used. The contact rates between farms were based on a study analyzing movement of pigs in Canada, while disease spread parameters were extracted from published literature. Eighteen distinct scenarios were designed and simulated by varying the mode of transmission (direct versus direct and indirect contact), type of index herd (farrowing, nursery and finishing), and the presumed network structures among swine farms (random, scale-free and small-world). PRRS virus was seeded in a randomly selected farm and 500 iterations of each scenario were simulated for 52 weeks. The median epidemic size by the end of the simulated period and percentage die-out for each scenario, were the key outcomes captured. Scenarios with scale-free network models resulted in the largest epidemic sizes, while scenarios with random and small-world network models resulted in smaller and similar epidemic sizes. Similarly, stochastic die-out percentage was least for scenarios with scale-free networks followed by random and small-world networks. Findings of the study indicated that incorporating network structures among the swine farms had a considerable impact on the spread of PRRS virus, highlighting the importance of understanding and incorporating realistic contact structures when developing infectious disease spread models for similar populations. PMID:26464321

  11. Aromatic Amino Acids in the Juxtamembrane Domain of Severe Acute Respiratory Syndrome Coronavirus Spike Glycoprotein Are Important for Receptor-Dependent Virus Entry and Cell-Cell Fusion?

    PubMed Central

    Howard, Megan W.; Travanty, Emily A.; Jeffers, Scott A.; Smith, M. K.; Wennier, Sonia T.; Thackray, Larissa B.; Holmes, Kathryn V.

    2008-01-01

    The severe acute respiratory syndrome coronavirus (SARS-CoV) spike glycoprotein (S) is a class I viral fusion protein that binds to its receptor glycoprotein, human angiotensin converting enzyme 2 (hACE2), and mediates virus entry and cell-cell fusion. The juxtamembrane domain (JMD) of S is an aromatic amino acid-rich region proximal to the transmembrane domain that is highly conserved in all coronaviruses. Alanine substitutions for one or two of the six aromatic residues in the JMD did not alter the surface expression of the SARS-CoV S proteins with a deletion of the C-terminal 19 amino acids (S ?19) or reduce binding to soluble human ACE2 (hACE2). However, hACE2-dependent entry of trypsin-treated retrovirus pseudotyped viruses expressing JMD mutant S ?19 proteins was greatly reduced. Single alanine substitutions for aromatic residues reduced entry to 10 to 60% of the wild-type level. The greatest reduction was caused by residues nearest the transmembrane domain. Four double alanine substitutions reduced entry to 5 to 10% of the wild-type level. Rapid hACE2-dependent S-mediated cell-cell fusion was reduced to 60 to 70% of the wild-type level for all single alanine substitutions and the Y1188A/Y1191A protein. S ?19 proteins with other double alanine substitutions reduced cell-cell fusion further, from 40% to less than 20% of wild-type levels. The aromatic amino acids in the JMD of the SARS-CoV S glycoprotein play critical roles in receptor-dependent virus-cell and cell-cell fusion. Because the JMD is so highly conserved in all coronavirus S proteins, it is a potential target for development of drugs that may inhibit virus entry and/or cell-cell fusion mediated by S proteins of all coronaviruses. PMID:18199653

  12. The early whole-blood transcriptional signature of dengue virus and features associated with progression to dengue shock syndrome in Vietnamese children and young adults.

    PubMed

    Hoang, Long Truong; Lynn, David J; Henn, Matt; Birren, Bruce W; Lennon, Niall J; Le, Phuong Thi; Duong, Kien Thi Hue; Nguyen, Tham Thi Hong; Mai, Lanh Ngoc; Farrar, Jeremy J; Hibberd, Martin L; Simmons, Cameron P

    2010-12-01

    Dengue is a pantropic public health problem. In children, dengue shock syndrome (DSS) is the most common life-threatening complication. The ability to predict which patients may develop DSS may improve triage and treatment. To this end, we conducted a nested case-control comparison of the early host transcriptional features in 24 DSS patients and 56 sex-, age-, and virus serotype-matched uncomplicated (UC) dengue patients. In the first instance, we defined the "early dengue" profile. The transcriptional signature in acute rather than convalescent samples (?72 h post-illness onset) was defined by an overabundance of interferon-inducible transcripts (31% of the 551 overabundant transcripts) and canonical gene ontology terms that included the following: response to virus, immune response, innate immune response, and inflammatory response. Pathway and network analyses identified STAT1, STAT2, STAT3, IRF7, IRF9, IRF1, CEBPB, and SP1 as key transcriptional factors mediating the early response. Strikingly, the only difference in the transcriptional signatures of early DSS and UC dengue cases was the greater abundance of several neutrophil-associated transcripts in patients who progressed to DSS, a finding supported by higher plasma concentrations of several canonical proteins associated with neutrophil degranulation (bactericidal/permeability-increasing protein [BPI], elastase 2 [ELA2], and defensin 1 alpha [DEF1A]). Elevated levels of neutrophil-associated transcripts were independent of the neutrophil count and also of the genotype of the infecting virus, as genome-length sequences of dengue virus serotype 1 (DENV-1) (n = 15) and DENV-2 (n = 3) sampled from DSS patients were phylogenetically indistinguishable from those sampled from uncomplicated dengue patients (32 DENV-1 and 9 DENV-2 sequences). Collectively, these data suggest a hitherto unrecognized association between neutrophil activation, pathogenesis, and the development of DSS and point to future strategies for guiding prognosis. PMID:20943967

  13. Lung pathogenicity of European genotype 3 strain porcine reproductive and respiratory syndrome virus (PRRSV) differs from that of subtype 1 strains.

    PubMed

    Weesendorp, Eefke; Rebel, Johanna M J; Popma-De Graaf, Ditta J; Fijten, Helmi P D; Stockhofe-Zurwieden, Norbert

    2014-11-01

    Porcine reproductive and respiratory syndrome (PRRS) is difficult to control due to a high mutation rate of the PRRS virus (PRRSV) and the emergence of virulent strains. The objective of this study was to analyse early and late pathological responses in the respiratory tract after infection with the European PRRSV subtype 3 strain Lena in comparison to two European PRRSV subtype 1 strains: Belgium A and Lelystad-Ter Huurne (LV). For each virus strain, groups of twelve pigs were inoculated, and four pigs per group were euthanized at days 3, 7 and 35 post-infection (p.i.) for consecutive examination. Infection with strain Lena resulted in a more severe disease than with the subtype 1 strains, an inflammatory response within the first week of infection with expression of IL-1? in the lung and lymph node, and an influx of neutrophils and monocytes in bronchoalveolar lavage fluid (BALF). Infection with strain Belgium A or LV resulted in mild or no pathology within the first week of infection, but inflammatory cell influx in the lung interstititium was increased at the end of the experiment at day 35 p.i. At five weeks p.i., all strains induced a higher percentage of cytotoxic T cells and higher levels of IFN-? producing cells in BALF. This might have contributed to clearance of virus. In general, subtype 3 strain Lena induced a stronger early inflammatory response which led to more severe clinical disease and pathology. On the other hand, this may have supported an enhanced or faster clearance of virus in tissues, compared to subtype 1 strains. PMID:25301281

  14. Cloning of profilin (FcPFN) from the shrimp Fenneropenaeus chinensis, a highly expressed protein in white spot syndrome virus (WSSV)-infected shrimp.

    PubMed

    Kong, H J; Hong, G-E; Cho, H K; Nam, B-H; Kim, Y-O; Kim, W-J; Lee, S-J; Kim, K-K

    2009-01-01

    We isolated and characterized the profilin (FcPFN) cDNA from hemocytes of Fenneropenaeus chinensis, a unique shrimp species from the Yellow Sea. The FcPFN cDNA consists of 830 bp and encodes a polypeptide of 125 amino acids, having a predicted isoelectric point of 5.06. The deduced amino acid sequence of FcPFN shows 36% and 90% amino acid sequence identity to the profilin genes of Pacific white shrimp Litopenaeus vannamei and black tiger shrimp Penaeus monodon, respectively. The FcPFN mRNA was highly expressed in hemocytes and hepatopancreas and moderately in muscle of normal shrimp. The higher expression of FcPFN mRNA is observed in shrimp infected with the white spot syndrome virus (WSSV), which is a major concern in all shrimp-growing regions of the world. These results suggest a potential role for FcPFN in viral host defense mechanisms. PMID:19638679

  15. Haptoglobin and C-reactive protein as biomarkers in the serum, saliva and meat juice of pigs experimentally infected with porcine reproductive and respiratory syndrome virus.

    PubMed

    Gómez-Laguna, Jaime; Gutiérrez, Ana; Pallarés, Francisco J; Salguero, Francisco J; Cerón, José J; Carrasco, Librado

    2010-07-01

    Concentrations of the acute phase proteins haptoglobin (Hp) and C-reactive protein (CRP) were measured in the serum, saliva and meat juice of pigs experimentally infected with a porcine reproductive and respiratory syndrome virus (PRRSv) field isolate. Sixteen PRRSv-free pigs were inoculated IM, killed in groups of four at 7, 14, 21 and 24 days post-inoculation (dpi), and samples of blood, saliva and diaphragmatic muscle were collected. Four non-infected controls were killed at 24 dpi. Significant differences in lung lesions were found between PRRSv-inoculated animals and controls. Changes in the concentrations of Hp and CRP in serum, saliva and meat juice samples were similar, peaking at 21 dpi. The correlations found suggest that the measurement of Hp and CRP in saliva and meat juice could serve as complementary, or possibly alternative, biomarkers of pig herd-health. PMID:20554227

  16. Full genome sequences of torque teno sus virus strains that coinfected a pig with postweaning multisystemic wasting syndrome in Japan: implications for genetic diversity.

    PubMed

    Ozawa, Makoto; Kawabata, Toshiko; Okuya, Kosuke; Nagano, Kiori; Kanda, Takehiro; Kanazawa, Norihiro; Tsukiyama-Kohara, Kyoko; Taneno, Akira; Deguchi, Eisaburo

    2015-12-01

    We determined the complete genome sequences of torque teno sus viruses (TTSuVs) detected in pigs with postweaning multisystemic wasting syndrome (PMWS) and in healthy pigs in Japan. Unexpectedly, we found coinfection of a PMWS-affected pig in Japan with one strain of TTSuV1, five strains of TTSuV2, and one strain of PCV2. Full-genome sequencing of each of these strains, followed by phylogenetic analysis, revealed broad genetic diversity in the TTSuV2 strains infecting the PMWS-affected pig. These results suggest that the geographical bias in the available genetic information about TTSuVs has a limited impact on the evaluation of their genetic diversity. PMID:26335893

  17. Application of a SYBR®Green one step real-time RT-PCR assay to detect type 1 porcine reproductive and respiratory syndrome virus.

    PubMed

    Iseki, Hiroshi; Takagi, Michihiro; Kuroda, Yoshiko; Katsuda, Ken; Mikami, Osamu; Tsunemitsu, Hiroshi; Yamakawa, Makoto

    2014-10-01

    The emergence in Japan of field isolates of type 1 porcine reproductive and respiratory syndrome virus (PRRSV) suggests problems with control. We therefore developed a one-step real-time reverse transcription polymerase chain reaction (qRT-PCR) with improved sensitivity that detects as little as 1 × 10(-2) TCID50/ml of viral RNA. We tested serum samples collected in January and September 2008, October 2009 and January 2011 from a farm with an outbreak and found infected pigs between January and September 2008, but not in January 2011. Further, between 2008 and 2011, we did not detect infection in pigs at 8 nearby farms or in 2,052 serum samples collected from pigs from 74 farms in 12 prefectures. This assay should help prevent future outbreaks. PMID:25047905

  18. Field and experimental data indicate that the eastern cottontail (Sylvilagus floridanus) is susceptible to infection with European brown hare syndrome (EBHS) virus and not with rabbit haemorrhagic disease (RHD) virus.

    PubMed

    Lavazza, Antonio; Cavadini, Patrizia; Barbieri, Ilaria; Tizzani, Paolo; Pinheiro, Ana; Abrantes, Joana; Esteves, Pedro J; Grilli, Guido; Gioia, Emanuela; Zanoni, Mariagrazia; Meneguz, Pier; Guitton, Jean-Sébastien; Marchandeau, Stéphane; Chiari, Mario; Capucci, Lorenzo

    2015-01-01

    The eastern cottontail (Sylvilagus floridanus) is an American lagomorph. In 1966, it was introduced to Italy, where it is currently widespread. Its ecological niche is similar to those of native rabbits and hares and increasing overlap in distribution brings these species into ever closer contact. Therefore, cottontails are at risk of infection with the two lagoviruses endemically present in Italy: Rabbit Haemorrhagic Disease virus (RHDV) and European Brown Hare Syndrome Virus (EBHSV). To verify the susceptibility of Sylvilagus to these viruses, we analyzed 471 sera and 108 individuals from cottontail populations in 9 provinces of north-central Italy from 1999 to 2012. In total, 15-20% of the cottontails tested seropositive for EBHSV; most titres were low, but some were as high as 1/1280. All the cottontails virologically tested for RHDV and EBHSV were negative with the exception of one individual found dead with hares during a natural EBHS outbreak in December 2009. The cottontail and the hares showed typical EBHS lesions, and the EBHSV strain identified was the same in both species (99.9% identity). To experimentally confirm the diagnosis, we performed two trials in which we infected cottontails with both EBHSV and RHDV. One out of four cottontails infected with EBHSV died of an EBHS-like disease, and the three surviving animals developed high EBHSV antibody titres. In contrast, neither mortality nor seroconversion was detected after infection with RHDV. Taken together, these results suggest that Sylvilagus is susceptible to EBHSV infection, which occasionally evolves to EBHS-like disease; the eastern cottontail could therefore be considered a "spill over" or "dead end" host for EBHSV unless further evidence is found to confirm that it plays an active role in the epidemiology of EBHSV. PMID:25828691

  19. Molecular Characterization of Transcriptome-wide Interactions between Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus and Porcine Alveolar Macrophages in vivo

    PubMed Central

    Zhou, Ping; Zhai, Shanli; Zhou, Xiang; Lin, Ping; Jiang, Tengfei; Hu, Xueying; Jiang, Yunbo; Wu, Bin; Zhang, Qingde; Xu, Xuewen; Li, Jin-ping; Liu, Bang

    2011-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) infects mainly the porcine alveolar macrophages (PAMs) and causes porcine reproductive and respiratory syndrome (PRRS). Previous studies have analyzed the global gene expression profiles of lung tissue in vivo and PAMs in vitro following infection with PRRSV, however, transcriptome-wide understanding of the interaction between highly pathogenic PRRSV (HP-PRRSV) and PAMs in vivo has not yet been established. In this study, we employed Affymetrix microarrays to investigate the gene expression patterns of PAMs isolated from Tongcheng piglets (a Chinese indigenous breed) after infection with HP-PRRSV. During the infection, Tongcheng piglets exhibited typical clinical signs, e.g. fever, asthma, coughing, anorexia, lethargy and convulsion, but displayed mild regional lung damage at 5 and 7 dpi. Microarray analysis revealed that HP-PRRSV infection has affected PAMs in expression of the important genes involved in cytoskeleton and exocytosis organization, protein degradation and folding, intracellular calcium and zinc homeostasis. Several potential antiviral strategies might be employed in PAMs, including upregulating IFN-induced genes and increasing intracellular zinc ion concentration. And inhibition of the complement system likely attenuated the lung damage during HP-PRRSV infection. Transcriptomic analysis of PAMs in vivo could lead to a better understanding of the HP-PRRSV-host interaction, and to the identification of novel antiviral therapies and genetic components of swine tolerance/susceptibility to HP-PRRS. PMID:21850204

  20. Identification of two auto-cleavage products of nonstructural protein 1 (nsp1) in porcine reproductive and respiratory syndrome virus infected cells: nsp1 function as interferon antagonist

    SciTech Connect

    Chen, Z.; Lawson, S.; Sun, Z.; Zhou, X.; Guan, X.; Christopher-Hennings, J.; Nelson, E.A.; Fang, Y.

    2010-03-01

    The porcine reproductive and respiratory syndrome virus nsp1 is predicted to be auto-cleaved from the replicase polyprotein into nsp1alpha and nsp1beta subunits. In infected cells, we detected the actual existence of nsp1alpha and nsp1beta. Cleavage sites between nsp1alpha/nsp1beta and nsp1beta/nsp2 were identified by protein microsequencing analysis. Time course study showed that nsp1alpha and nsp1beta mainly localize into the cell nucleus after 10 h post infection. Further analysis revealed that both proteins dramatically inhibited IFN-beta expression. The nsp1beta was observed to significantly inhibit expression from an interferon-stimulated response element promoter after Sendai virus infection or interferon treatment. It was further determined to inhibit nuclear translocation of STAT1 in the JAK-STAT signaling pathway. These results demonstrated that nsp1beta has ability to inhibit both interferon synthesis and signaling, while nsp1alpha alone strongly inhibits interferon synthesis. These findings provide important insights into mechanisms of nsp1 in PRRSV pathogenesis and its impact in vaccine development.

  1. Identification of CD8+ cytotoxic T lymphocyte epitopes from porcine reproductive and respiratory syndrome virus matrix protein in BALB/c mice

    PubMed Central

    2011-01-01

    Twenty-seven nanopeptides derived from the matrix (M) protein of porcine reproductive and respiratory syndrome virus (PRRSV) were screened for their ability to elicit a recall interferon-? (IFN-?) response from the splenocytes of BALB/c mice following DNA vaccination and a booster vaccination with recombinant vaccinia virus rWR-PRRSV-M. We identified two peptides (amino acid residues K93FITSRCRL and F57GYMTFVHF) as CD8+ cytotoxic T lymphocyte (CTL) epitopes. These peptides elicited significant numbers of IFN-? secreting cells, compared with other M nonapeptides and one irrelevant nonapeptide. Bioinformatics analysis showed that the former is an H-2Kd-restricted CTL epitope, and the latter is an H-2Dd-restricted CTL epitope. Multiple amino acid sequence alignment among different PRRSV M sequences submitted to GenBank indicated that these two CTL epitopes are strongly conserved, and they should therefore be considered for further research on the mechanisms of cellular immune responses to PRRSV. PMID:21619712

  2. Association of Epstein-Barr virus (EBV) with Sjögren's syndrome: differential EBV expression between epithelial cells and lymphocytes in salivary glands.

    PubMed Central

    Wen, S.; Shimizu, N.; Yoshiyama, H.; Mizugaki, Y.; Shinozaki, F.; Takada, K.

    1996-01-01

    The association of Epstein Barr virus (EBV) with Sjögren's syndrome (SS) is still in dispute. This study is aimed to investigate the existence of EBV genomes and their products in salivary glands of SS. Salivary gland samples were surgically obtained from Chinese patients. EBV DNA was detected in three of seven cases by dot blot hybridization and in four of seven cases by in situ hybridization. The EBV-encoded small RNA-1 (EBER1) was detected in two of seven cases by in situ hybridization. The immunohistochemical staining of EBV proteins showed that the EBV latent membrane protein-1 was detected in four of seven cases and that BZLF1, BALF2, and gp350/220 proteins associating with virus production were not expressed. In eight controls, no positive signal was observed by these methods. DNA in situ hybridization identified ERV on both epithelial cells and lymphocytes. On the other hand, EBER1-positive signals were exclusively localized on lymphocytes. These results indicate that two forms of EBV infection may exist in salivary glands of SS. One is EBER1-positive latency in lymphocytes, the other is EBER1-negative latency in epithelial cells. Frequent EBV detection in salivary glands of SS suggests that EBV plays a role in the genesis of SS. Images Figure 1 Figure 2 Figure 3 PMID:8909241

  3. A one-step RT-PCR assay to detect and discriminate porcine reproductive and respiratory syndrome viruses in clinical specimens.

    PubMed

    Yang, Keli; Li, Yanhe; Duan, Zhengying; Guo, Rui; Liu, Zewen; Zhou, Danna; Yuan, Fangyan; Tian, Yongxiang

    2013-12-01

    Outbreaks of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) have led to large economic losses and, subsequently, have drawn great attention to its diagnosis and prevention. To facilitate rapid discrimination of HP-PRRSV from classical PRRSV (C-PRRSV), we developed a one-step RT-PCR assay. Primer specificities were evaluated with RNA extracted from 8 viral strains and our results revealed that the primers had a high specificity for PRRSV. The assay sensitivity was 25 copies/?L for both HP-PRRSV and C-PRRSV. A total of 929 serum samples were identified, of which 20.45% were HP-PRRSV-positive and 1.51% were C-PRRSV-positive, which was completely consistent with that of immunochromatochemistry and sequencing method. The proposed assay can detect the virus 2 days prior the onset of symptoms and it can be performed in 2h, thereby providing a rapid method to discriminate HP-PRRSV from C-PRRSV for the identification and prevention of PRRSV infections. PMID:24035936

  4. Expression of Toll-like receptor signaling-related genes in pigs co-infected with porcine reproductive and respiratory syndrome virus and porcine circovirus type 2.

    PubMed

    Dong, Van Hieu; Tu, Pang-Yan; Tsai, Pei-Chun; Lin, Yi-Hsin; Chang, Hsiu-Luan; Kuo, Tsun-Yung; Chiou, Ming-Tang; Lin, Chao-Nan; Chung, Wen-Bin

    2015-08-01

    Pigs co-infected with porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) have been shown to develop more severe diseases than pigs infected with PRRSV or PCV2 only. The underlying interaction mechanisms between the two viruses in developing the disease are unclear. The present study investigates the mRNA expression of Toll-like receptor (TLR) signaling-related molecules in peripheral blood mononuclear cells from pigs infected with PRRSV or PCV2 or both. The mRNA expression levels were determined by quantitative real-time RT-PCR. Co-infection of pigs with PRRSV and PCV2 resulted in a negatively synergistic effect on the mRNA expression of the negative regulators of TLR, including A20, Bcl-3, IRAK-M, MKP-1, SARM1 and SIGIRR, as well as the TLR downstream transcription factors IRF-1 and IRF-3. A positively synergistic effect of a combined infection of PRRSV and PCV2 on the CD14 mRNA expression was also observed. PMID:26022073

  5. Rapid and sensitive detection of type II porcine reproductive and respiratory syndrome virus by reverse transcription loop-mediated isothermal amplification combined with a vertical flow visualization strip.

    PubMed

    Gou, Hongchao; Deng, Jieru; Pei, Jingjing; Wang, Jiaying; Liu, Wenjun; Zhao, Mingqiu; Chen, Jinding

    2014-12-01

    Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was combined with a vertical flow (VF) nucleic acid detection strip to develop a universal assay for the detection of type II porcine reproductive and respiratory syndrome virus (PRRSV). The loop primers were labeled separately with biotin and fluorescein isothiocyanate (FITC) in this assay. Using optimized parameters, the whole reaction could be completed in <50 min in a completely enclosed environment. The detection limit of this assay was found to be 1 pg RNA, 30 tissue culture infective dose 50 (TCID50) virus, or 230 copies of recombinant plasmid DNA, which is relatively higher than that of RT-LAMP analyzed by agarose gel, RT-LAMP visualized by calcein, and the conventional RT-polymerase chain reaction (PCR). No false-positive results were obtained in the specificity assay. The efficiency of the RT-LAMP method was tested by analyzing 43 clinical samples, and the results were compared with those obtained by RT-PCR analysis, with the respective positive rates of 32.56% and 27.91%. This result confirmed that the method described is a rapid, accurate, and sensitive method for universal type II PRRSV detection. Also, this method can be used for the rapid detection of type II PRRSV during the early phase of an outbreak, especially for rapid veterinary diagnosis on the spot and in rural areas. PMID:25241142

  6. The application of a duplex reverse transcription real-time PCR for the surveillance of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2.

    PubMed

    Chang, Chia-Yi; Deng, Ming-Chung; Wang, Fun-In; Tsai, Hsiang-Jung; Yang, Chia-Huei; Chang, Chieh; Huang, Yu-Liang

    2014-06-01

    The porcine respiratory disease complex (PRDC) is the most common disease in commercial pork production worldwide. Porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), the most important agents of PRDC, usually co-infect in the same pigs. In order to survey the prevalence of PCV2 and PRRSV in pigs of various ages, a duplex reverse transcription real-time PCR (DRT-rPCR) was developed and applied in the present study. The DRT-rPCR did not cross-react with 10 swine viruses other than PCV2 and PRRSV, with detection limits of 1 TCID50/ml for PCV2 and 6.3 TCID50/ml for PRRSV. Surveillance using DRT-rPCR together with serology revealed that in the five farms studied, pigs were most susceptible to PRRSV at 6-14 weeks of age, whereas susceptibility to PCV2 varied by the management system but was mostly at 10-14 weeks of age. Cross analysis of viral loads versus antibody titers revealed that PCV2 load was affected negatively by anti-PCV2 ORF2 antibody, which constituted the most important non-infectious factor affecting the development of PMWS. These results indicated that DRT-rPCR was developed and applied successfully to the surveillance of PCV2 and PRRSV in the field. PMID:24560782

  7. Complete genomic characterization and genetic diversity of four European genotype porcine reproductive and respiratory syndrome virus isolates from China in 2011.

    PubMed

    Zhou, Zhi; Liu, Qi; Hu, Dongmei; Zhang, Qian; Han, Tao; Ma, Ying; Gu, Xiaoxue; Zhai, Xinyan; Tian, Kegong

    2015-12-01

    Porcine reproductive and respiratory syndrome (PRRS) is an emerging disease that has caused serious economic losses to the swine industry worldwide. In 2011, a nation-wide surveillance program investigated the prevalence of PRRS viruses (PRRSV) in Chinese breeding swine farms, and four European genotype PRRSV (PRRSV-Type 1) were successfully isolated. To explore the genetic diversity of PRRSV-Type 1 in China, these 4 viral strains were subjected to genome sequencing and analysis. The four isolates shared 87.4-90.7 % nucleotide homology with the Lelystad strain (PRRSV-Type 1 stereotype strain). NSP2, ORF3, and ORF4 were the most variable regions and contained discontinuous deletions or insertions when compared to other PRRSV-Type 1 strains. All isolates fell into separate branches of the subtype 1 of PRRSV-Type 1 phylogenetic tree. This analysis of emerging PRRSV-Type 1 strains revealed previously unrecorded genetic diversity. Close attention should be paid to the prevention and control of this evolving virus. PMID:26573283

  8. Crystal Structures of Major Envelope Proteins VP26 and VP28 from White Spot Syndrome Virus Shed Light on Their Evolutionary Relationship

    SciTech Connect

    Tang,X.; Wu, J.; Sivaraman, J.; Hew, C.

    2007-01-01

    White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be the first molecules to interact with the host. Here, we report the localization and crystal structure of major envelope proteins VP26 and VP28 from WSSV at resolutions of 2.2 and 2.0 {angstrom}, respectively. These two proteins alone account for approximately 60% of the envelope, and their structures represent the first two structural envelope proteins of WSSV. Structural comparisons among VP26, VP28, and other viral proteins reveal an evolutionary relationship between WSSV envelope proteins and structural proteins from other viruses. Both proteins adopt {beta}-barrel architecture with a protruding N-terminal region. We have investigated the localization of VP26 and VP28 using immunoelectron microscopy. This study suggests that VP26 and VP28 are located on the outer surface of the virus and are observed as a surface protrusion in the WSSV envelope, and this is the first convincing observation for VP26. Based on our studies combined with the literature, we speculate that the predicted N-terminal transmembrane region of VP26 and VP28 may anchor on the viral envelope membrane, making the core {beta}-barrel protrude outside the envelope, possibly to interact with the host receptor or to fuse with the host cell membrane for effective transfer of the viral infection. Furthermore, it is tempting to extend this host interaction mode to other structural viral proteins of similar structures. Our finding has the potential to extend further toward drug and vaccine development against WSSV.

  9. Porcine reproductive and respiratory syndrome virus (PRRSV) surveillance using pre-weaning oral fluid samples detects circulation of wild-type PRRSV.

    PubMed

    Kittawornrat, Apisit; Panyasing, Yaowalak; Goodell, Christa; Wang, Chong; Gauger, Phillip; Harmon, Karen; Rauh, Rolf; Desfresne, Luc; Levis, Ian; Zimmerman, Jeffrey

    2014-01-31

    Oral fluid samples collected from litters of piglets (n=600) one day prior to weaning were evaluated as a method to surveil for porcine reproductive and respiratory syndrome virus (PRRSV) infections in four sow herds of approximately 12,500 sow each. Serum samples from the litters' dam (n=600) were included for comparison. All four herds were endemically infected with PRRSV and all sows had been vaccinated ? 2 times with PRRSV modified-live virus vaccines. After all specimens had been collected, samples were randomized and assayed by PRRSV real-time reverse transcription polymerase chain reaction (RT-qPCR) and four PRRSV antibody ELISA assays (IgM, IgA, IgG, and Commercial Kit). All sow serum samples were negative by PRRSV RT-qPCR, but 9 of 600 oral fluid samples tested positive at two laboratories. Open reading frame 5 (ORF5) sequencing of 2 of the 9 positive oral fluid samples identified wild-type viruses as the source of the infection. A comparison of antibody responses in RT-qPCR positive vs. negative oral fluid samples showed significantly higher IgG S/P ratios in RT-qPCR-positive oral fluid samples (mean S/P 3.46 vs. 2.36; p=0.02). Likewise, sow serum samples from RT-qPCR-positive litter oral fluid samples showed significantly higher serum IgG (mean S/P 1.73 vs. 0.98; p<0.001) and Commercial Kit (mean S/P 1.97 vs. 0.98; p<0.001) S/P ratios. Overall, the study showed that pre-weaning litter oral fluid samples could provide an efficient and sensitive approach to surveil for PRRSV in infected, vaccinated, or presumed-negative pig breeding herds. PMID:24393634

  10. Development and validation of an assay to detect porcine reproductive and respiratory syndrome virus-specific neutralizing antibody titers in pig oral fluid samples.

    PubMed

    Ouyang, Kang; Binjawadagi, Basavaraj; Kittawornrat, Apisit; Olsen, Chris; Hiremath, Jagadish; Elkalifa, Nadia; Schleappi, Rose; Wu, Jianmin; Zimmerman, Jeffrey; Renukaradhya, Gourapura J

    2013-08-01

    Porcine reproductive and respiratory syndrome virus (PRRSV)-specific neutralizing antibodies (NA) are important for clearing the virus. Pen-based pig oral fluid samples for disease surveillance are gaining in importance due to the ease of collection and low cost. The aim of this study was to develop a PRRSV-specific NA assay to determine NA titers in pig oral fluid samples. At first, we standardized the PRRSV NA assay using pen-based pig oral fluid samples collected over a period of 3 months from a herd of swine that received a PRRSV modified live vaccine (PRRS-MLV), and we also used oral fluid and serum samples collected from individual boars that were vaccinated with PRRS-MLV or infected with a virulent PRRSV strain. Our results suggest that a PRRSV NA titer of >8 in oral fluid samples is virus specific and can be detected beginning at 28 days after vaccination or infection. To validate the assay, we used 104 pen-based pig oral fluid and five representative serum samples from each pen of unknown history, as well as 100 serum samples from repeatedly vaccinated sows and oral fluid samples of their respective litters belonging to four different swine-breeding farms. Our results demonstrated that PRRSV NA titers in oral fluid samples are correlated with serum sample titers, and maternally derived PRRSV-specific NA titers could be detected in litters at the time of weaning. In conclusion, we have standardized and validated the pig oral fluid-based PRRSV NA assay, which has 94.3% specificity and 90.5% repeatability. The assay can be used to monitor herd immunity against PRRSV in vaccinated and infected herds of swine. PMID:23784856

  11. A serological survey on classical swine fever (CSF), Aujeszky's disease (AD) and porcine reproductive and respiratory syndrome (PRRS) virus infections in French wild boars from 1991 to 1998.

    PubMed

    Albina, E; Mesplède, A; Chenut, G; Le Potier, M F; Bourbao, G; Le Gal, S; Leforban, Y

    2000-11-15

    In early 1992, a CSF epizootic was clinically recognised in a wild boar population of approximately 1300 animals within an area of 250km(2) located in the east of France. In order to check the CSF situation in wild boars outside this area, a serological survey was carried out in the rest of France, for 8 consecutive years (1991-1998). This paper reports on the results obtained during this survey which included wild boars shot during the hunting period but also boars reared within fences. Around 1000-2700 sera a year were tested for the presence of antibodies to classical swine fever virus (CSFV) and also to Aujeszky's disease virus (ADV). Out of 12025 sera tested over the whole period, 80 wild boars were found positive for CSF antibodies. Sixty of them were collected on wild boars shot during the years 1992-1994 in the epizootic area located in east of France and 10 were collected in Corsica during the years 1994-1996. The last four positive samples were single reactors coming from areas or farms, which were thereafter confirmed to be serologically negative. These results together with the fact that no disease has been reported so far illustrate that the French wild boar population is probably not concerned by CSF infection (excepted in the east of France where the disease has now become enzootic). Two hundred and forty nine sera were initially detected as CSF positive but confirmed secondarily as positive for border disease (BD) antibodies. This finding shows that wild boars are also susceptible to infection by ruminant pestiviruses. Four hundred and twenty three wild boars have been found positive for ADV antibodies. In addition, from 1993 to 1995, 909 samples were tested for the presence of antibodies to porcine reproductive and respiratory syndrome virus (PRRSV). Thirty three of them were positive. The results on AD and PRRS antibody detection show that wild boars may constitute a reservoir for various infectious diseases of pigs. PMID:11042399

  12. Recombinant Modified Vaccinia Virus Ankara Expressing the Spike Glycoprotein of Severe Acute Respiratory Syndrome Coronavirus Induces Protective Neutralizing Antibodies Primarily Targeting the Receptor Binding Region

    PubMed Central

    Chen, Zhiwei; Zhang, Linqi; Qin, Chuan; Ba, Lei; Yi, Christopher E.; Zhang, Fengwen; Wei, Qiang; He, Tian; Yu, Wenjie; Yu, Jian; Gao, Hong; Tu, Xinming; Gettie, Agegnehu; Farzan, Michael; Yuen, Kwok-yung; Ho, David D.

    2005-01-01

    Immunization with a killed or inactivated viral vaccine provides significant protection in animals against challenge with certain corresponding pathogenic coronaviruses (CoVs). However, the promise of this approach in humans is hampered by serious concerns over the risk of leaking live severe acute respiratory syndrome (SARS) viruses. In this study, we generated a SARS vaccine candidate by using the live-attenuated modified vaccinia virus Ankara (MVA) as a vector. The full-length SARS-CoV envelope Spike (S) glycoprotein gene was introduced into the deletion III region of the MVA genome. The newly generated recombinant MVA, ADS-MVA, is replication incompetent in mammalian cells and highly immunogenic in terms of inducing potent neutralizing antibodies in mice, rabbits, and monkeys. After two intramuscular vaccinations with ADS-MVA alone, the 50% inhibitory concentration in serum was achieved with reciprocal sera dilutions of more than 1,000- to 10,000-fold in these animals. Using fragmented S genes as immunogens, we also mapped a neutralizing epitope in the region of N-terminal 400 to 600 amino acids of the S glycoprotein (S400-600), which overlaps with the angiotensin-converting enzyme 2 (ACE2) receptor-binding region (RBR; S318-510). Moreover, using a recombinant soluble RBR-Fc protein, we were able to absorb and remove the majority of the neutralizing antibodies despite observing that the full S protein tends to induce a broader spectrum of neutralizing activities in comparison with fragmented S proteins. Our data suggest that a major mechanism for neutralizing SARS-CoV likely occurs through blocking the interaction between virus and the cellular receptor ACE2. In addition, ADS-MVA induced potent immune responses which very likely protected Chinese rhesus monkeys from pathogenic SARS-CoV challenge. PMID:15708987

  13. High production of the acquired immunodeficiency syndrome virus (lymphadenopathy-associated virus) by human T lymphocytes stimulated by streptococcal mitogenic toxins.

    PubMed Central

    Alouf, J E; Geoffroy, C; Klatzmann, D; Gluckman, J C; Gruest, J; Montagnier, L

    1986-01-01

    Purified streptococcal mitogens (SMs) including erythrogenic exotoxin were compared with phytohemagglutinin (PHA) for their ability to sustain lymphadenopathy-associated virus (LAV) replication after the stimulation of normal human peripheral blood mononuclear cells and purified CD4+ and CD8+ T cells infected with LAV. Both SM and PHA supported LAV production in peripheral blood mononuclear and CD4+ cells but not in CD8+ cells. LAV production assessed by the assay of reverse transcriptase in cell supernatants appeared earlier after stimulation with SM and was 6- to 10-fold greater than after stimulation by PHA. PMID:3490491

  14. 50 CFR 648.105 - Possession restrictions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... with or possess nets or pieces of net on board that do not meet the minimum mesh requirements and that... through April 30, unless the vessel possesses a valid summer flounder small-mesh exemption LOA and...

  15. [Virus--the causative agent of hemorrhagic fever with renal syndrome--a new representative of the family Bunyaviridae].

    PubMed

    Donets, M A; Rezapkin, G V; Korolev, M B; Tkachenko, E A

    1983-01-01

    Physicochemical characteristics (sedimentation coefficient, buoyant density of particles, as well as ribonuclein of these particles) and morphology of the causative agent of hemorrhagic fever with renal syndrome were found to be similar to those of the other members of the Bunyaviridae family. PMID:6412459

  16. Severe acute respiratory syndrome (SARS)

    MedlinePLUS

    Severe acute respiratory syndrome (SARS) is a serious form of pneumonia . It is caused by a virus that was first identified ... chap 366. McIntosh K, Perlman S. Coronaviruses, including severe ... syndrome (SARS)and Middle East respiratory syndrome (MERS). In: ...

  17. Genomic analysis and pathogenic characteristics of Type 2 porcine reproductive and respiratory syndrome virus nsp2 deletion strains isolated in Korea.

    PubMed

    Choi, Hwan-Won; Nam, Eeuri; Lee, Yoo Jin; Noh, Yun-Hee; Lee, Seung-Chul; Yoon, In-Joong; Kim, Hyun-Soo; Kang, Shien-Young; Choi, Young-Ki; Lee, Changhee

    2014-06-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) is a globally ubiquitous swine virus that exhibits genetic and pathogenic heterogeneity among isolates. The present study was conducted to determine the complete genome sequence and pathogenicity of two Korean type 2 PRRSV nonstructural protein 2 (nsp2) deletion mutants, CA-2 and KNU-12-KJ4. The full-length genomes of CA-2 and KNU-12-KJ4 were determined to be 15,018 and 15,019 nucleotides in length, excluding the poly(A) tail, respectively, which were 393- or 392-nucleotide shorter than that of the type 2 NA prototype strain VR-2332 due to the presence of notable large deletions within the nsp2 gene. The genomes of CA-2 and KNU-12-KJ4 consisted of a 189- or 190-nucleotide 5' untranslated region (UTR), a 14,677-nucleotide protein-coding region, and a 151-nucleotide 3' UTR. Whole genome evaluation revealed that the nucleotide sequences of CA-2 and KNU-12-KJ4 are most similar to each other (10.7% sequence divergence), and then to the Korean strain CA-1 (11.3% sequence divergence) and the US strain MN184C (13.1% sequence divergence), respectively. To evaluate the in vitro immunity of nsp2 deletion variants, we sought to explore alteration of inflammatory cytokine and chemokine expression in PAM-pCD163 cells infected with each virus strain using quantitative real-time RT-PCR. Cytokine genes including IL-8, IL-10, and TNF-?, and chemokines such as MCP-1 and RANTES were found to be significantly elevated in nsp2 deletion virus-infected PAM cells. In contrast, expression of interferons (IFN-?, ?, and ?) and antiviral genes including ISG-15, -54, and -56 were unchanged or down-regulated in PAM cells infected with the nsp2 deletion mutants. Animal studies to assess the pathogenicity of nsp2 deletion PRRSVs demonstrated that both CA-2 and KNU-12-KJ4 strains notably produce weight loss in infected pigs. Furthermore, the nsp2 deletion mutants replicated well in pigs with significantly increased and prolonged viremia kinetics. Taken together, our results indicate that, among the three isolates, the outcome of in vitro and in vivo infection by CA-2 and KNU-12-KJ4 is comparable, suggesting that the large nsp2 deletion may be one of the viral genetic determinants contributing to PRRSV pathogenicity. PMID:24646599

  18. Tumorigenic DNA viruses

    SciTech Connect

    Klein, G.

    1989-01-01

    The eighth volume of Advances in Viral Oncology focuses on the three major DNA virus groups with a postulated or proven tumorigenic potential: papillomaviruses, animal hepatitis viruses, and the Epstein-Bar virus. In the opening chapters, the contributors analyze the evidence that papillomaviruses and animal hepatitis viruses are involved in tumorigenesis and describe the mechanisms that trigger virus-host cell interactions. A detailed section on the Epstein-Barr virus (EBV) - comprising more than half the book - examines the transcription and mRNA processing patterns of the virus genome; the mechanisms by which EBV infects lymphoid and epithelial cells; the immunological aspects of the virus; the actions of EBV in hosts with Acquired Immune Deficiency Syndrome; and the involvement of EBV in the etiology of Burkitt's lymphoma.

  19. The human oncogenic viruses

    SciTech Connect

    Luderer, A.A.; Weetall, H.H

    1986-01-01

    This book contains eight selections. The titles are: Cytogenetics of the Leukemias and Lymphomas; Cytogenetics of Solid Tumors: Renal Cell Carcinoma, Malignant Melanoma, Retinoblastoma, and Wilms' Tumor; Elucidation of a Normal Function for a Human Proto-Oncogene; Detection of HSV-2 Genes and Gene Products in Cervical Neoplasia; Papillomaviruses in Anogennital Neoplasms; Human Epstein-Barr Virus and Cancer; Hepatitis B Virus and Hepatocellular Carcinoma; and Kaposi's Sarcoma: Acquired Immunodeficiency Syndrome (AIDS) and Associated Viruses.

  20. Phylogenetic comparison of porcine circovirus type 2 (PCV2) and porcine reproductive respiratory syndrome virus (PRRSV) strains detected in domestic pigs until 2008 and in 2012 in Croatia

    PubMed Central

    2014-01-01

    Background Porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) have been present for the last 2 decades in Croatia, causing large economical losses in the pig production. The clinical features of the infections are mostly manifested by the development of respiratory problems, weight loss and poor growth performance, as well as reproductive failure in pregnant sows. Even though the infections are continuously recognized in some regions in Croatia, the heterogeneity of the detected viral strains from 2012 has not yet been investigated. The objective of this study was to compare virus strains of PCV2 and PRRSV detected until 2008 in Croatia with strains isolated in 2012 to gain a better epidemiological understanding of these two infections. Results PCV2 and PRRSV strains detected in 2012 in fattening pigs from regions where these two diseases have been previously described were compared to strains that have been detected in the same regions within the past two decades. The phylogenetic analysis revealed that the circulating PCV2 and PRRSV strains are distantly related to the previously described Croatian viral strains. However, when compared to known isolates from the GenBank a high genetic identity of PRRSV isolates with isolates from Hungary, Denmark and the Netherlands was found. Conclusion The results of this study reveal that even though PCV2 and PRRSV are constantly present in the investigated regions in Croatia, the viral strains found in 2012 genetically differ from those detected in earlier years. This indicates that new entries into the pig population appeared with regard to both infections, probably as a result of pig trade. PMID:24839544

  1. An evaluation of disinfectants for the sanitation of porcine reproductive and respiratory syndrome virus-contaminated transport vehicles at cold temperatures.

    PubMed

    Dee, Scott; Deen, John; Burns, Danny; Douthit, George; Pijoan, Carlos

    2005-01-01

    The objective of this study was to evaluate the efficacy of commercially available disinfectants to sanitize porcine reproductive and respiratory syndrome virus (PRRSV) contaminated trailer models in cold climates (-20 degrees C and 4 degrees C). Disinfectants evaluated included Synergize, Aseptol 2000, Biophene, Sentramax, Virkon, Tek Trol, and DC&R. All products were applied to trailers via fumigation at 4 degrees C. Following experimental contamination of model trailers with PRRSV MN 30-100 (5 x 10(5) TCID50), models were tested for the presence or absence of PRRSV-RNA by polymerase chain reaction (PCR) on swabs collected 0, 30, and 60 min after treatment. Treatments included washing only, washing plus disinfectant fumigation, washing plus fumigation, and washing plus overnight drying. The PRRSV-RNA detected across trailers ranged from 0/12 replicates in trailers treated with Synergize or allowed to dry for 8 h. These trailers were also negative for the presence of infectious PRRSV, based on the lack of sentinel pig infection (0/4 replicates). In contrast, the detection of PRRSV-positive swabs by PCR ranged from 3/12 (Aseptol) to 10/12 (Biophene). Based on these results, the efficacy of Synergize was evaluated at -20 degrees C. In an attempt to reduce the impact of freezing on disinfectant activity, 30 mL of disinfectant was added to a 3840 mL of a 40% methanol solution, a 10% propylene glycol (PG) solution, or water alone. The PRRSV-contaminated trailers were treated with 1 of 3 disinfectant mixtures via fumigation, stored for 8 h at -20 degrees C, allowed to thaw, and sampled as described. Trailers treated with 40% methanol or 10% PG did not freeze and were negative for PRRSV-RNA and infectious virus following thawing. In contrast, trailers treated with disinfectant and water were frozen within 60 min at -20 degrees C, and decontamination was not successful. PMID:15745225

  2. Probability of detecting Porcine reproductive and respiratory syndrome virus infection using pen-based swine oral fluid specimens as a function of within-pen prevalence.

    PubMed

    Olsen, Chris; Wang, Chong; Christopher-Hennings, Jane; Doolittle, Kent; Harmon, Karen M; Abate, Sarah; Kittawornrat, Apisit; Lizano, Sergio; Main, Rodger; Nelson, Eric A; Otterson, Tracy; Panyasing, Yaowalak; Rademacher, Chris; Rauh, Rolf; Shah, Rohan; Zimmerman, Jeffrey

    2013-05-01

    Pen-based oral fluid sampling has proven to be an efficient method for surveillance of infectious diseases in swine populations. To better interpret diagnostic results, the performance of oral fluid assays (antibody- and nucleic acid-based) must be established for pen-based oral fluid samples. Therefore, the objective of the current study was to determine the probability of detecting Porcine reproductive and respiratory syndrome virus (PRRSV) infection in pen-based oral fluid samples from pens of known PRRSV prevalence. In 1 commercial swine barn, 25 pens were assigned to 1 of 5 levels of PRRSV prevalence (0%, 4%, 12%, 20%, or 36%) by placing a fixed number (0, 1, 3, 5, or 9) of PRRSV-positive pigs (14 days post PRRSV modified live virus vaccination) in each pen. Prior to placement of the vaccinated pigs, 1 oral fluid sample was collected from each pen. Thereafter, 5 oral fluid samples were collected from each pen, for a total of 150 samples. To confirm individual pig PRRSV status, serum samples from the PRRSV-negative pigs (n = 535) and the PRRSV vaccinated pigs (n = 90) were tested for PRRSV antibodies and PRRSV RNA. The 150 pen-based oral fluid samples were assayed for PRRSV antibody and PRRSV RNA at 6 laboratories. Among the 100 samples from pens containing ?1 positive pig (?4% prevalence) and tested at the 6 laboratories, the mean positivity was 62% for PRRSV RNA and 61% for PRRSV antibody. These results support the use of pen-based oral fluid sampling for PRRSV surveillance in commercial pig populations. PMID:23536612

  3. Characterization of immune responses following homologous reinfection of pigs with European subtype 1 and 3 porcine reproductive and respiratory syndrome virus strains that differ in virulence.

    PubMed

    Weesendorp, Eefke; Stockhofe-Zurwieden, Norbert; Nauwynck, Hans J; Popma-De Graaf, Ditta J; Rebel, Johanna M J

    2016-01-15

    Porcine reproductive and respiratory syndrome virus (PRRSV) causes significant economic losses to the pork industry worldwide. Vaccination results often in limited protection. Understanding host immune responses elicited by different PRRSV strains could help to develop more efficacious vaccines. In the current study we characterized immunological responses and viral kinetics in pigs after primo infection and homologous challenge of the highly virulent European subtype 3 strain Lena, and the moderate to low virulent subtype 1 strain LV. Eighteen pigs were infected per strain, and 18 non-infected pigs served as control. Post mortem analysis was performed at days 7, 46 and 60 p.i. At day 46, pigs were challenged with the homologous strain. After the first inoculation, pigs infected with Lena developed fever and clinical symptoms, while this was not observed in pigs infected with LV. Virus titres in serum were about 100-fold higher in pigs infected with Lena than in pigs infected with LV. An inflammatory response was observed in pigs after primo infection with Lena with significantly higher levels of IL-12, IL-1? and TNF-? in the bronchoalveolar lavage. IFN-? ELISPOT assay showed comparable responses between Lena and LV. Neutralizing antibodies were detected earlier in serum of pigs infected with Lena than in pigs infected with LV. After the challenge, a boost in antibody levels in both groups was observed. Challenge infection resulted in both groups in complete protection and sterile immunity, with no viraemia, clinical symptoms or viral RNA in tissues. In conclusion, although there were clear differences in immunological, clinical and virological responses to the primo infection, there were no differences observed in protection against homologous challenge. PMID:26711030

  4. The pathogenicity determinant of Citrus tristeza virus causing the seedling yellows syndrome maps at the 3'-terminal region of the viral genome.

    PubMed

    Albiach-Marti, Maria R; Robertson, Cecile; Gowda, Siddarame; Tatineni, Satyanarayana; Belliure, Belén; Garnsey, Stephen M; Folimonova, Svetlana Y; Moreno, Pedro; Dawson, William O

    2010-01-01

    Citrus tristeza virus (CTV) (genus Closterovirus, family Closteroviridae) causes some of the more important viral diseases of citrus worldwide. The ability to map disease-inducing determinants of CTV is needed to develop better diagnostic and disease control procedures. A distinctive phenotype of some isolates of CTV is the ability to induce seedling yellows (SY) in sour orange, lemon and grapefruit seedlings. In Florida, the decline isolate of CTV, T36, induces SY, whereas a widely distributed mild isolate, T30, does not. To delimit the viral sequences associated with the SY syndrome, we created a number of T36/T30 hybrids by substituting T30 sequences into different regions of the 3' half of the genome of an infectious cDNA of T36. Eleven T36/T30 hybrids replicated in Nicotiana benthamiana protoplasts. Five of these hybrids formed viable virions that were mechanically transmitted to Citrus macrophylla, a permissive host for CTV. All induced systemic infections, similar to that of the parental T36 clone. Tissues from these C. macrophylla source plants were then used to graft inoculate sour orange and grapefruit seedlings. Inoculation with three of the T30/T36 hybrid constructs induced SY symptoms identical to those of T36; however, two hybrids with T30 substitutions in the p23-3' nontranslated region (NTR) (nucleotides 18 394-19 296) failed to induce SY. Sour orange seedlings infected with a recombinant non-SY p23-3' NTR hybrid also remained symptomless when challenged with the parental virus (T36), demonstrating the potential feasibility of using engineered constructs of CTV to mitigate disease. PMID:20078776

  5. Positive inductive effect of swine interleukin-4 on immune responses elicited by modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine.

    PubMed

    Peng, Jun; Wang, Jinbao; Wu, Jiaqiang; Du, Yijun; Li, Jun; Guo, Zhongkun; Yu, Jiang; Xu, Shaojian; Zhang, Yuyu; Sun, Wenbo; Cong, Xiaoyan; Shi, Jianli

    2013-12-01

    Porcine reproductive and respiratory syndrome (PRRS) has become one of the most economically important diseases to the global pork industry. Currently, the efficacies of available commercial vaccines remain questionable: the modified live-PRRSV vaccines (MLVs) were generally effective but variable in sufficient protection, and the outcomes of inactivated-PRRSV vaccines (IVs) in the field were not very promising. In the present study, we investigated the effect of swine interleukin 4 (IL-4) on the development of virus-specific immune responses elicited by an MLV. The antibody titer against PRRSV membrane proteins in pigs elicited by MLV plus recombinant plasmid encoding IL-4 (group 3) was significantly higher than those elicited by MLV alone (group 1) and MLV plus empty plasmid (group 2) from 35 days post-inoculation (dpi). Similarly, the neutralizing efficacy of sera from group 3 was markedly enhanced compared with group 1 and group 2. In cellular immunity, the ratio of CD3?CD4?/CD3?CD8? T lymphocyte subpopulations from group 3 monitored by flow cytometry (FCM) was significantly higher than those from group 1 and group 2 from 42?dpi to 21 days post-challenge (dpc). After viral challenge, pigs in group 3 showed significantly lower virus loads in peripheral blood measured by a real-time quantitative PCR (RT-qPCR), as compared with those in group 1 and group 2. Pigs in group 1 and group 2 had a low fever and displayed mild inappetence, lethargy, rough hair coats, and no lung lesions, while those in group 3 showed almost no clinical signs, no lung lesions. The scores of clinical signs of pigs in group 3 were significantly lower than those in both group 1 and group 2. Interestingly, the scores of lung lesions showed no significant differences among the three groups. Our results indicate that swine IL-4 markedly enhanced the protective immune response of pigs and improved the efficacy of the MLV in preventing PRRS disease. PMID:24328935

  6. Glycosyl-phosphatidylinositol (GPI)-anchored membrane association of the porcine reproductive and respiratory syndrome virus GP4 glycoprotein and its co-localization with CD163 in lipid rafts

    SciTech Connect

    Du, Yijun; Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan ; Pattnaik, Asit K.; Song, Cheng; Yoo, Dongwan; Li, Gang; Institute of Animal Science and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing

    2012-03-01

    The porcine reproductive and respiratory syndrome virus (PRRSV) glycoprotein 4 (GP4) resembles a typical type I membrane protein in its structure but lacks a hydrophilic tail at the C-terminus, suggesting that GP4 may be a lipid-anchored membrane protein. Using the human decay-accelerating factor (DAF; CD55), a known glycosyl-phosphatidylinositol (GPI) lipid-anchored protein, chimeric constructs were made to substitute the GPI-anchor domain of DAF with the putative lipid-anchor domain of GP4, and their membrane association and lipase cleavage were determined in cells. The DAF-GP4 fusion protein was transported to the plasma membrane and was cleaved by phosphatidylinositol-specific phospholipase C (PI-PLC), indicating that the C-terminal domain of GP4 functions as a GPI anchor. Mutational studies for residues adjacent to the GPI modification site and characterization of respective mutant viruses generated from infectious cDNA clones show that the ability of GP4 for membrane association corresponded to virus viability and growth characteristics. The residues T158 ({omega} - 2, where {omega} is the GPI moiety at E160), P159 ({omega} - 1), and M162 ({omega} + 2) of GP4 were determined to be important for virus replication, with M162 being of particular importance for virus infectivity. The complete removal of the peptide-anchor domain in GP4 resulted in a complete loss of virus infectivity. The depletion of cholesterol from the plasma membrane of cells reduced the virus production, suggesting a role of lipid rafts in PRRSV infection. Remarkably, GP4 was found to co-localize with CD163 in the lipid rafts on the plasma membrane. Since CD163 has been reported as a cellular receptor for PRRSV and GP4 has been shown to interact with this receptor, our data implicates an important role of lipid rafts during entry of the virus.

  7. Replication characteristics of eight virulent and two attenuated genotype 1 and 2 porcine reproductive and respiratory syndrome virus (PRRSV) strains in nasal mucosa explants.

    PubMed

    Frydas, Ilias S; Nauwynck, Hans J

    2016-01-15

    Porcine reproductive and respiratory syndrome virus (PRRSV) can spread in between pigs via contact and airborne route. It was shown before that the highly pathogenic PRRSV strain Lena was able to replicate 10-100 times more in the nasal mucosa compared to the low pathogenic PRRSV strain LV. In this work, the replication characteristics of four type 1 (LV, 07V063, 08VA, 13V091), three type 2 (VR2332, MN-184, VN) and two attenuated (MLV-DV, MLV-VR2332) PRRSV strains were studied. After 72hpi, mean virus titers reached 10(4.5-4.8) TCID50/ml for LV and 08VA, 10(5.2-5.4) TCID50/ml for VR2332 and Lena, and 10(5.8-6.3) TCID50/ml for 07V063, 13V091, MN-184 and VN strains, whereas attenuated strains remained below detection limit. The mean number of PRRSV-positive cells/mm(2) at 72hpi was 1.1 and 1.3 for the attenuated strains and LV, 13.3 for 08VA, 23.5 and 29.3 for VR2332 and 07063, 31.1 and 33.8 for 13V091 and Lena, and, 39.1 and 59.2 for MN-184 and VN respectively. All the LV and MLV-LV infected cells were Sn(+), whereas all other strains also infected Sn(-) macrophages. In conclusion, (i) based on the virus shedding in the respiratory explants, PRRSV strains can be categorized as poor (MLV-DV, MLV-VR2332, LV, 08VA), moderate (Lena, VR2332) and strong (07V063, 13V091, MN-184, VN) secretors, and (ii) based on the number of infected cells isolates can be categorized as low (MLV-DV, MLV-VR2332, LV), moderately (08VA, VR2332), highly (07V063, Lena, 13V091) and hyper (MN-184, VN) virulent in the nasal mucosa. PMID:26711043

  8. Porcine reproductive and respiratory syndrome virus infection is associated with an increased number of Sn-positive and CD8-positive cells in the maternal-fetal interface.

    PubMed

    Karniychuk, U U; De Spiegelaere, W; Nauwynck, H J

    2013-09-01

    It is already known that porcine reproductive and respiratory syndrome virus (PRRSV) infection in lungs changes a local cell pattern and cytokine profile. However, there is no information about cellular and immunological events upon PRRSV infection in the maternal-fetal interface yet. The altered number and/or function of macrophages and NK cells in the maternal-fetal interface during infection may have a functional importance for virus replication. In addition, local cellular and immunological disbalance may also disrupt fragile homeostasis and contribute to the PRRSV-related reproductive disorders. Sialoadhesin (Sn)-positive macrophages are target cells for PRRSV and Sn overexpression has been observed upon chronic inflammatory and infectious diseases. It is also known that mouse Sn-positive macrophages in lymph nodes are able to closely interact with and activate NK cells in response to viral particles. Therefore, the main purpose of the present study was to examine if PRRSV infection is associated with altered Sn expression on endometrial and placental macrophages. In addition, CD8-positive cells (porcine endometrial NK cells were previously described as CD8(+)CD3(-) cells) were localized and quantified in the PRRSV-positive and control tissues. Tissue samples were obtained from three PRRSV-inoculated and three non-inoculated control sows at 100 days of gestation. Real-time RT-PCR showed a clear upregulation of Sn mRNA expression in the PRRSV-positive endometrium/placenta (p<0.05). Sn-, CD163- and CD14-specific immunofluorescence stainings revealed that PRRSV-inoculated sows had a significantly higher number of Sn-positive macrophages in the endometrium and placenta due to de novo Sn expression on local CD163-positive macrophages. Along with the increased number of Sn-positive macrophages an increased number of CD8-positive cells, which were mostly CD3-negative, was observed in the PRRSV-positive endometrium. The effects of the observed cellular changes on virus replication and potential contribution to placental damage and reproductive disorders are discussed. PMID:23707347

  9. Ocular Tropism of Respiratory Viruses

    PubMed Central

    Rota, Paul A.; Tumpey, Terrence M.

    2013-01-01

    SUMMARY Respiratory viruses (including adenovirus, influenza virus, respiratory syncytial virus, coronavirus, and rhinovirus) cause a broad spectrum of disease in humans, ranging from mild influenza-like symptoms to acute respiratory failure. While species D adenoviruses and subtype H7 influenza viruses are known to possess an ocular tropism, documented human ocular disease has been reported following infection with all principal respiratory viruses. In this review, we describe the anatomical proximity and cellular receptor distribution between ocular and respiratory tissues. All major respiratory viruses and their association with human ocular disease are discussed. Research utilizing in vitro and in vivo models to study the ability of respiratory viruses to use the eye as a portal of entry as well as a primary site of virus replication is highlighted. Identification of shared receptor-binding preferences, host responses, and laboratory modeling protocols among these viruses provides a needed bridge between clinical and laboratory studies of virus tropism. PMID:23471620

  10. Vaccination with a Porcine Reproductive and Respiratory Syndrome (PRRS) Modified Live Virus Vaccine Followed by Challenge with PRRS Virus and Porcine Circovirus Type 2 (PCV2) Protects against PRRS but Enhances PCV2 Replication and Pathogenesis Compared to Results for Nonvaccinated Cochallenged Controls.

    PubMed

    Niederwerder, Megan C; Bawa, Bhupinder; Serão, Nick V L; Trible, Benjamin R; Kerrigan, Maureen A; Lunney, Joan K; Dekkers, Jack C M; Rowland, Raymond R R

    2015-12-01

    Coinfections involving porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) contribute to a group of disease syndromes known as porcine circovirus-associated disease (PCVAD). Presumably, PRRSV infection enhances PCV2 replication as a result of modulation of host immunity. The purpose of this study was to evaluate PCV2 replication and pathogenesis in pigs vaccinated with a PRRS modified live virus (MLV) vaccine and subsequently challenged with a combination of PRRSV and PCV2. During the early postchallenge period, the number of pigs with PRRSV-associated clinical signs was decreased, and average daily gain (ADG) was increased, in the vaccinated group, demonstrating the protective effect of PRRS vaccination. However, during the later postchallenge period, more pigs in the vaccinated group showed increased PCV2 viremia, decreased ADG, increased PCVAD clinical signs, and increased mortality. In this disease model, the early benefits of PRRSV vaccination were outweighed by the later amplification of PCVAD. PMID:26446422

  11. The Correlation Between Perceived Social Support and Illness Uncertainty in People with Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome in Iran

    PubMed Central

    Sajjadi, Moosa; Rassouli, Maryam; Bahri, Narges; Mohammadipoor, Fatemeh

    2015-01-01

    Background: Illness uncertainty is a source of a chronic and pervasive psychological stress for people living with human immunodeficiency virus (HIV)/acquired immune deficiency syndrome (AIDS) (PLWH), and largely affects their quality of life and the ability to cope with the disease. Based on the uncertainty in illness theory, the social support is one of the illness uncertainty antecedents, and influences the level of uncertainty perceived by patients. Aim: To examine uncertainty in PLWH and its correlation with social support in Iran. Materials and Methods: This cross-sectional correlational study was conducted with 80 PLWH presenting to AIDS Research Center, Tehran, Iran in 2013. The data collected using illness uncertainty and social support inventories were analyzed through Pearson's correlation coefficient, Spearman's correlation coefficient, and regression analysis. Results: The results showed a high level of illness uncertainty in PLWH and a negative significant correlation between perceived social support and illness uncertainty (P = 0.01, r = -0.29). Conclusion: Uncertainty is a serious aspect of illness experience in Iranian PLWH. Providing adequate, structured information to patients as well as opportunities to discuss their concerns with other PLWH and receive emotional support from their health care providers may be worthwhile. PMID:26009679

  12. Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp

    SciTech Connect

    Wang Yuzhen; Zhang Xiaohua; Yuan Li; Xu Tao; Rao Yu; Li Jia; Dai Heping

    2008-08-08

    White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron microscopy. Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV.

  13. Consensus on context-specific strategies for reducing the stigma of human immunodeficiency virus/acquired immunodeficiency syndrome in Zambézia Province, Mozambique

    PubMed Central

    Mukolo, Abraham; Torres, Isabel; Bechtel, Ruth M.; Sidat, Mohsin; Vergara, Alfredo E.

    2014-01-01

    Stigma has been implicated in poor outcomes of human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) care. Reducing stigma is important for HIV prevention and long-term treatment success. Although stigma reduction interventions are conducted in Mozambique, little is known about the current nature of stigma and the efficacy and effectiveness of stigma reduction initiatives. We describe action research to generate consensus on critical characteristics of HIV stigma and anti-stigma interventions in Zambézia Province, Mozambique. Qualitative data gathering methods, including indepth key-informant interviews, community interviews and consensus group sessions, were utilized. Delphi methods and the strategic options development analysis technique were used to synthesize qualitative data. Key findings are that stigma enacted by the general public might be declining in tandem with the HIV/AIDS epidemic in Mozambique, but there is likely excessive residual fear of HIV disease and community attitudes that sustain high levels of perceived stigma. HIV-positive women accessing maternal and child health services appear to shoulder a disproportionate burden of stigma. Unintentional biases among healthcare providers are currently the critical frontier of stigmatization, but there are few interventions designed to address them. Culturally sensitive psychotherapies are needed to address psychological distress associated with internalized stigma and these interventions should complement current supports for voluntary counseling and testing. While advantageous for defining stakeholder priorities for stigma reduction efforts, confirmatory quantitative studies of these consensus positions are needed before the launch of specific interventions. PMID:24527744

  14. Identification of a Novel Nonstructural Protein, VP9, from White Spot Syndrome Virus: Its Structure Reveals a Ferredoxin Fold with Specific Metal Binding Sites

    SciTech Connect

    Liu,Y.; Wu, J.; Song, J.; Sivaraman, J.; Hew, C.

    2006-01-01

    White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP9, a full-length protein of WSSV, encoded by open reading frame wsv230, was identified for the first time in the infected Penaeus monodon shrimp tissues, gill, and stomach as a novel, nonstructural protein by Western blotting, mass spectrometry, and immunoelectron microscopy. Real-time reverse transcription-PCR demonstrated that the transcription of VP9 started from the early to the late stage of WSSV infection as a major mRNA species. The structure of full-length VP9 was determined by both X-ray and nuclear magnetic resonance (NMR) techniques. It is the first structure to be reported for WSSV proteins. The crystal structure of VP9 revealed a ferredoxin fold with divalent metal ion binding sites. Cadmium sulfate was found to be essential for crystallization. The Cd2+ ions were bound between the monomer interfaces of the homodimer. Various divalent metal ions have been titrated against VP9, and their interactions were analyzed using NMR spectroscopy. The titration data indicated that VP9 binds with both Zn2+ and Cd2+. VP9 adopts a similar fold as the DNA binding domain of the papillomavirus E2 protein. Based on our present investigations, we hypothesize that VP9 might be involved in the transcriptional regulation of WSSV, a function similar to that of the E2 protein during papillomavirus infection of the host cells.

  15. The 15N and 46R Residues of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein Enhance Regulatory T Lymphocytes Proliferation

    PubMed Central

    Bai, Juan; Li, Yufeng; Zhang, Qiaoya; Jiang, Ping

    2015-01-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) negatively modulates host immune responses, resulting in persistent infection and immunosuppression. PRRSV infection increases the number of PRRSV-specific regulatory T lymphocytes (Tregs) in infected pigs. However, the target antigens for Tregs proliferation in PRRSV infection have not been fully understood. In this study, we demonstrated that the highly pathogenic PRRSV (HP-PRRSV) induced more CD4+CD25+Foxp3+ Tregs than classical PRRSV (C-PRRSV) strain. Of the recombinant GP5, M and N proteins of HP-PRRSV expressed in baculovirus expression systems, only N protein induced Tregs proliferation. The Tregs assays showed that three amino-acid regions, 15–21, 42–48 and 88–94, in N protein played an important role in induction of Tregs proliferation with synthetic peptides covering the whole length of N protein. By using reverse genetic methods, it was firstly found that the 15N and 46R residues in PRRSV N protein were critical for induction of Tregs proliferation. The phenotype of induced Tregs closely resembled that of transforming-growth-factor-?-secreting T helper 3 Tregs in swine. These data should be useful for understanding the mechanism of immunity to PRRSV and development of infection control strategies in the future. PMID:26397116

  16. Is human immunodeficiency virus/acquired immunodeficiency syndrome decreasing among Brazilian injection drug users? Recent findings and how to interpret them.

    PubMed

    Bastos, Francisco I; Bongertz, Vera; Teixeira, Sylvia Lopes; Morgado, Mariza G; Hacker, Mariana A

    2005-02-01

    We briefly review findings from Brazilian settings where the human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) epidemic among injection drug users (IDUs) seems to be decreasing, highlighting recent findings from Rio de Janeiro and discussing methodological alternatives. Former analyses using serologic testing algorithm for recent HIV seroconversion have shown that HIV incidence has been low in IDUs recruited by two different surveys carried out in Rio, where low injection frequencies and infection rates have been found among new injectors. The proportion of AIDS cases among IDUs in Rio has been fairly modest, compared to São Paulo and especially to the southernmost states. Notwithstanding, the interpretation of findings from serial surveys constitutes a challenge, magnified in the assessment of HIV spread among IDUs due to the dynamic nature of the drug scenes and limitations of sampling strategies targeting hard-to-reach populations. Assessment of epidemic trends may profit from the triangulation of data, but cannot avert biases associated with sampling errors. Efforts should be made to triangulate data from different sources, besides exploring specific studies from different perspectives. In an attempt to further assess the observed trends, we carried out original analyses using data from Brazilian AIDS databank. PMID:15867971

  17. Porcine reproductive and respiratory syndrome virus (PRRSV) in serum and oral fluid samples from individual boars: will oral fluid replace serum for PRRSV surveillance?

    PubMed

    Kittawornrat, Apisit; Prickett, John; Chittick, Wayne; Wang, Chong; Engle, Mark; Johnson, Jeremy; Patnayak, Devi; Schwartz, Trevor; Whitney, Daniel; Olsen, Chris; Schwartz, Kent; Zimmerman, Jeffrey

    2010-12-01

    The purpose of this study was to determine whether oral fluid samples could be used to monitor individually-housed adult boars for porcine reproductive and respiratory syndrome virus (PRRSV) infection. In 3 trials, 24 boars were intramuscularly (IM) inoculated with a modified-live PRRSV (MLV) vaccine (Trial 1), a Type 1 PRRSV isolate (Trial 2), or a Type 2 isolate (Trial 3). Oral fluid samples were collected daily and serum samples were collected twice weekly. Following the completion of the study, samples were randomized and blind-tested for PRRSV by real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR). PRRSV was detected in oral fluids at DPI 1 and all oral fluid specimens were PRRSV qRT-PCR positive at DPI 4. Although PRRSV was detected in both serum and oral fluid specimens through DPI 21, a comparison of matched samples from individual boars showed that oral fluid was equal to serum for the detection of PRRSV at DPI 7 and more likely to be positive than serum on DPI 14 and 21. Overall, oral fluid was superior to serum for the detection of PRRSV using PCR over the 21-day observation period in this study. The results of this experiment suggest that individually-penned oral fluid sampling could be an efficient, cost-effective approach to PRRSV surveillance in boar studs and other swine populations. PMID:20670665

  18. Effect of saliva stabilisers on detection of porcine reproductive and respiratory syndrome virus in oral fluid by quantitative reverse transcriptase real-time PCR.

    PubMed

    Decorte, Inge; Van der Stede, Yves; Nauwynck, Hans; De Regge, Nick; Cay, Ann Brigitte

    2013-08-01

    This study evaluated the effect of extraction-amplification methods, storage temperature and saliva stabilisers on detection of porcine reproductive and respiratory syndrome virus (PRRSV) RNA by quantitative reverse transcriptase real-time PCR (qRT-PCR) in porcine oral fluid. The diagnostic performance of different extraction-amplification methods was examined using a dilution series of oral fluid spiked with PRRSV. To determine RNA stability, porcine oral fluid, with or without commercially available saliva stabilisers, was spiked with PRRSV, stored at 4°C or room temperature and tested for the presence of PRRSV RNA by qRT-PCR. PRRSV RNA could be detected in oral fluid using all extraction-amplification combinations, but the limit of detection varied amongst different combinations. Storage temperature and saliva stabilisers had an effect on the stability of PRRSV RNA, which could only be detected for 7 days when PRRSV spiked oral fluid was kept at 4°C or stabilised at room temperature with a commercial mRNA stabiliser. PMID:23489844

  19. Downregulation of antigen-presenting cells in tonsil and lymph nodes of porcine reproductive and respiratory syndrome virus-infected pigs.

    PubMed

    Rodríguez-Gómez, I M; Gómez-Laguna, J; Barranco, I; Pallarés, F J; Ramis, G; Salguero, F J; Carrasco, L

    2013-10-01

    Porcine reproductive and respiratory syndrome virus (PRRSV) can persist in different organs of infected pigs, which suggests a failure in the immune response. Antigen-presenting cells (APCs) play a pivotal role in the induction of effective T- and B-cell responses. In this study, we investigated the changes in the different APC subpopulations and T- and B-cell counts in the tonsil, retropharyngeal and mediastinal lymph nodes of pigs experimentally infected with a European PRRSV field isolate. Our results demonstrated that the expression of S100, SWC3, HLA-DR molecule and CD3 was diminished in the studied organs throughout the study, observing a significant negative correlation between viral antigen and HLA-DR expression in both retropharyngeal and mediastinal lymph nodes. In contrast, ?-light chains showed an increase during the study. Taking all into account, after PRRSV infection, no enhancement in the number of APCs and T cells was observed, suggesting an impairment of the immune function which may allow the persistence of PRRSV into the organism. PMID:22816521

  20. Taura syndrome virus IRES initiates translation by binding its tRNA-mRNA–like structural element in the ribosomal decoding center

    PubMed Central

    Koh, Cha San; Brilot, Axel F.; Grigorieff, Nikolaus; Korostelev, Andrei A.

    2014-01-01

    In cap-dependent translation initiation, the open reading frame (ORF) of mRNA is established by the placement of the AUG start codon and initiator tRNA in the ribosomal peptidyl (P) site. Internal ribosome entry sites (IRESs) promote translation of mRNAs in a cap-independent manner. We report two structures of the ribosome-bound Taura syndrome virus (TSV) IRES belonging to the family of Dicistroviridae intergenic IRESs. Intersubunit rotational states differ in these structures, suggesting that ribosome dynamics play a role in IRES translocation. Pseudoknot I of the IRES occupies the ribosomal decoding center at the aminoacyl (A) site in a manner resembling that of the tRNA anticodon-mRNA codon. The structures reveal that the TSV IRES initiates translation by a previously unseen mechanism, which is conceptually distinct from initiator tRNA-dependent mechanisms. Specifically, the ORF of the IRES-driven mRNA is established by the placement of the preceding tRNA-mRNA–like structure in the A site, whereas the 40S P site remains unoccupied during this initial step. PMID:24927574

  1. Type 2 Porcine Reproductive and Respiratory Syndrome Virus infection mediated apoptosis in B- and T-cell areas in lymphoid organs of experimentally infected pigs.

    PubMed

    Gómez-Laguna, J; Salguero, F J; Fernández de Marco, M; Barranco, I; Rodríguez-Gómez, I M; Quezada, M; Carrasco, L

    2013-06-01

    Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection is characterized by persisting in lungs and lymphoid tissue, resulting in systemic lymphoid depletion. The aim of this study was to correlate the histological changes, viral antigen expression and apoptosis phenomena in tonsil, medial retropharyngeal and mediastinal lymph nodes of 12 pigs inoculated with a type 2 PRRSV isolate (Chilean strain 2402). Apoptosis phenomena were observed mainly in lymphocytes and secondly in macrophages of lymph nodes and tonsils of inoculated animals, showing a peak of both apoptotic cells and viral antigen expression at the end of the study (21 dpi). However, the number of apoptotic cells was higher than the number of PRRSV-positive cells at the end of the study. This finding together with the location of apoptotic cells and PRRSV-positive cells in different structures of lymphoid organs supports the hypothesis that PRRSV-positive macrophages might modulate the apoptosis phenomena in other cells, mainly lymphocytes, by means of an indirect mechanism. Furthermore, apoptotic cells were detected both in B- and T-cell areas of lymphoid organs, suggesting that apoptosis phenomena may play a role in the impairment of the host immune response during PRRS. PMID:22607093

  2. Porcine reproductive and respiratory syndrome virus: interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods.

    PubMed

    Wernike, Kerstin; Bonilauri, Paolo; Dauber, Malte; Errington, Jane; LeBlanc, Neil; Revilla-Fernández, Sandra; Hjulsager, Charlotte; Isaksson, Mats; Stadejek, Tomasz; Beer, Martin; Hoffmann, Bernd

    2012-09-01

    To compare the real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays used for the diagnosis of Porcine reproductive and respiratory syndrome virus (PRRSV), a Europe-wide interlaboratory ring trial was conducted. A variety of PRRSV strains including North American (NA) and European (EU) genotype isolates were analyzed by the participants. Great differences regarding qualitative diagnostics as well as analytical sensitivity were observed between the individual RT-qPCR systems, especially when investigating strains from the EU genotype. None of the assays or commercial kits used in the ring trial could identify all different PRRSV strains with an optimal analytical and diagnostic sensitivity. The genetic variability of the PRRSV strains, which is supposed to hinder the diagnostic of the RT-PCR because of mutations at the primer binding sites, was also confirmed by sequencing and subsequent phylogenetic analysis. In summary, a major problem in PRRSV diagnostics by RT-qPCR is false-negative results. To achieve maximum safety in the molecular diagnosis of PRRSV, the combined usage of different assays or kits is highly recommended. PMID:22807507

  3. Expression profile of Toll-like receptor mRNA in pigs co-infected with porcine reproductive and respiratory syndrome virus and porcine circovirus type 2.

    PubMed

    Tu, Pang-Yan; Tsai, Pei-Chun; Lin, Yi-Hsin; Liu, Po-Cheng; Chang, Hsiu-luan; Kuo, Tsun-Yung; Chung, Wen-Bin

    2015-02-01

    Field and experimental studies have shown that co-infection of pigs with porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) increases the severity of the disease. The present study investigates the mRNA expression profile of Toll-like receptors (TLRs) in pigs co-infected with PRRSV and PCV2. SPF pigs were infected with PRRSV, PCV2 or in a combination of both. The mRNA expression levels of TLRs and related cytokines in peripheral blood mononuclear cells (PBMCs) of pigs were determined by quantitative real-time RT-PCR. The mRNA expression profiles of TLRs by PBMCs from pigs co-infected with PRRSV and PCV2 displayed two distinct patterns: an increased expression profile for TLRs2, 4 and 8, and a decreased expression profile for TLRs3, 7 and 9. An up-regulated expression of IL-1? and IL-10 mRNA and a down-regulated expression of INF-? and TNF-? mRNA in PBMCs of co-infected pigs were also observed. PMID:25555603

  4. Cellular microRNA miR-26a suppresses replication of porcine reproductive and respiratory syndrome virus by activating innate antiviral immunity

    PubMed Central

    Jia, Xiaojuan; Bi, Yuhai; Li, Jing; Xie, Qing; Yang, Hanchun; Liu, Wenjun

    2015-01-01

    Porcine reproductive and respiratory syndrome (PRRS) has caused large economic losses in the swine industry in recent years. Current PRRS vaccines fail to effectively prevent and control this disease. Consequently, there is a need to develop new antiviral strategies. MicroRNAs play critical roles in intricate host-pathogen interaction networks, but the involvement of miRNAs during PRRS virus (PRRSV) infection is not well understood. In this study, pretreatment with miR-26a induced a significant inhibition of PRRSV replication and remission of the cytopathic effect in MARC-145 cells, and this antiviral effect was sustained for at least 120?h. Luciferase reporter analysis showed that the PRRSV genome was not the target of miRNA-26a. Instead, RNA-seq analysis demonstrated that miR-26a significantly up-regulated innate anti-viral responses, including activating the type I interferon (IFN) signaling pathway and promoting the production of IFN-stimulated genes. These findings suggest that delivery of miR-26a may provide a potential strategy for anti-PRRSV therapies. PMID:26013676

  5. 50 CFR 648.125 - Possession limit.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... possession limit. The owner, operator, and crew of a charter or party boat issued a scup moratorium permit... crew members for a party boat, or more than three crew members for a charter boat. This possession.... (d) Scup and scup parts harvested by a vessel with a moratorium or charter or party boat scup...

  6. Epilepsy and religious experiences: Voodoo possession.

    PubMed

    Carrazana, E; DeToledo, J; Tatum, W; Rivas-Vasquez, R; Rey, G; Wheeler, S

    1999-02-01

    Epileptic seizures have a historical association with religion, primarily through the concept of spirit possession. Five cases where epileptic seizures were initially attributed to Voodoo spirit possession are presented. The attribution is discussed within the context of the Voodoo belief system. PMID:9952273

  7. 50 CFR 648.105 - Possession restrictions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... § 648.4(a)(3) that fish with or possess nets or pieces of net on board that do not meet the minimum mesh... November 1 through April 30, unless the vessel possesses a valid summer flounder small-mesh exemption...

  8. Jugular foramen syndrome caused by varicella zoster virus infection in a patient with ipsilateral hypoplasia of the jugular foramen.

    PubMed

    Hayashi, T; Murayama, S; Sakurai, M; Kanazawa, I

    2000-01-01

    We report a patient with acute cranial polyneuropathy with unilateral involvement of the ninth, tenth, and eleventh cranial nerves. Although this patient lacked a typical cutaneous herpetic manifestation, elevated levels of IgM and IgG antibodies to varicella zoster virus (VZV) in both the serum and cerebrospinal fluid confirmed the clinical diagnosis of VZV infection and zoster sine herpete. Coexisting hypoplasia of the ipsilateral jugular foramen was detected using three-dimensional, surface-rendering displays reconstructed from the cranial helical CT scan. The patient recovered almost completely following treatment with an anti-inflammatory corticosteroid. Anatomical narrowing of the jugular foramen in this patient may have contributed to entrapment of the affected nerves at their passage through the foramen. PMID:10620663

  9. Expression of antigenic epitopes of porcine reproductive and respiratory syndrome virus (PRRSV) in a modified live-attenuated porcine circovirus type 2 (PCV2) vaccine virus (PCV1-2a) as a potential bivalent vaccine against both PCV2 and PRRSV.

    PubMed

    Piñeyro, Pablo E; Kenney, Scott P; Giménez-Lirola, Luis G; Heffron, C Lynn; Matzinger, Shannon R; Opriessnig, Tanja; Meng, Xiang-Jin

    2015-12-01

    Co-infection of pigs in the field with porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) is common and poses a major concern in effective control of PCV2 and PRRSV. We previously demonstrated that insertion of foreign epitope tags in the C-terminus of PCV2 ORF2 produced infectious virions that elicited humoral immune responses against both PCV2 capsid and inserted epitope tags. In this study, we aimed to determine whether the non-pathogenic chimeric virus PCV1-2a, which is the basis for the licensed PCV2 vaccine Fostera™ PCV, can express PRRSV antigenic epitopes, thus generating dual immunity as a potential bivalent vaccine against both PCV2 and PPRSV. Four different linear B-cell antigenic epitopes of PRRSV were inserted into the C-terminus of the capsid gene of the PCV1-2a vaccine virus. We showed that insertion of 12 (PRRSV-GP2 epitope II, PRRSV-GP3 epitope I, and PRRSV-GP5 epitope I), and 14 (PRRSV-GP5 epitope IV) amino acid residues did not impair the replication of the resulting PCV1-2a-PRRSVEPI chimeric viruses in vitro. The four chimeric PCV1-2a viruses expressing PRRSV B-cell linear epitopes were successfully rescued and characterized. An immunogenicity study in pigs revealed that two of the four chimeric viruses, PCV1-2a-PRRSVEPIGP3IG and PCV1-2a-PRRSVEPIEPIGP5IV, elicited neutralizing antibodies against PRRSV VR2385 as well as PCV2 (strains PCV2a, PCV2b, and mPCV2b). The results have important implications for exploring the potential use of PCV1-2a vaccine virus as a live virus vector to develop bivalent MLVs against both PCV2 and PRRSV. PMID:26239318

  10. Porcine reproductive and respiratory syndrome virus detection in Thailand during 2005-2010 in relation to clinical problems, pig types, regions, and seasons.

    PubMed

    Tummaruk, Padet; Surapat, Pannin; Sriariyakun, Sutharat; Seemakram, Oraphan; Olanratmanee, Em-on; Tantilertcharoen, Rachod; Thanawongnuwech, Roongroj

    2013-03-01

    The objectives of the present study were to determine the prevalence of porcine reproductive and respiratory syndrome virus (PRRSV) in Thailand between 2005 and 2010. The study was conducted by retrospectively investigating the detection of PRRSV from different pig types including boars, sows, piglets, nursery pigs, and fattening pigs from six regions of Thailand, i.e., the northern, eastern, northeastern, central, western, and southern parts. The data were obtained from cases submitted to the Chulalongkorn University Veterinary Diagnostic Laboratory for PRRSV detection between 2005 and 2010. Frequency analyses and generalized linear models were used to evaluate the prevalence of PRRSV in relation to various factors. In total, 2,273 tissues (n?=?636), semen (n?=?210) and serum (n?=?1,427) samples were included. PRRSV was detected in 32.6 % (740/2,273) of the pigs. The virus was found in 43.1 %, 15.7 %, and 30.3 % in the tissues, semen, and serum samples, respectively (P?

  11. Regulation of toll-like receptors 3, 7 and 9 in porcine alveolar macrophages by different genotype 1 strains of porcine reproductive and respiratory syndrome virus.

    PubMed

    Kuzemtseva, Liudmila; de la Torre, Eugenia; Martín, Gerard; Soldevila, Ferran; Ait-Ali, Tahar; Mateu, Enric; Darwich, Laila

    2014-04-15

    The toll-like receptors (TLRs) play an important role in the innate host defense against pathogens. Endosomal TLRs, TLR3, TLR7/8, and TLR9 are involved in antiviral responses by promoting the production of antiviral cytokines such as type I interferons. Porcine reproductive and respiratory syndrome (PRRS) is an important disease causing economically high losses to the swine industry worldwide and caused by a single stranded positive sense RNA virus, known as PRRS virus (PRRSV). Studies focused on the interaction between PRRSV and TLRs are scarce. The aim of the present study was to evaluate the expression of TLR3, TLR7 and TLR9 in porcine alveolar macrophages (PAM) infected with different genotype 1 PRRSV strains previously sequenced and characterized by their ability to induce TNF-?: 3262 (TNF-? inducer), 3267 (TNF-? not inducer) and an attenuated vaccine strain (strain Deventer, PorcilisPRRS, Merck) that replicates scarcely in PAM. PAM were infected with the different PRRSV strains (at 0.1 multiplicity of infection) for 48 h or mock-stimulated with PAM supernatants. Cells were collected at different time-points (0 h, 6 h, 12 h, 24 h, 36 h, 48 h) to determine the kinetics of viral replication by quantitative RT-PCR (qRT-PCR) and the expression of TLR3, 7 and 9 by qRT-PCR, flow cytometry and indirect immunofluorescence assay. Although infection with PRRSV did not affect significantly relative levels of any TLR mRNA transcript (normalized to ?-actin expression), this infection resulted in significant differences in the proportion of cells expressing TLR3. Thus, in PAM infected with PRRSV strain 3262 the proportion of TLR3+ cells significantly increased from 24h compared with the controls; in contrast strain 3267 resulted in a lower proportion of TLR3+ PAM. Interestingly, strain 3262 replicate to lower levels than 3267 at comparable post-inoculation times. For strain DV, the results indicated that this strain did not replicate substantially in PAM and did not stimulated TLR3 expression. These observations suggest that at least TLR3 is regulated differentially by different genotype 1 PRRSV strains and this seems to be related apparently to the replication levels of each strain, as well as, to the TNF-? inducing capability. The fact that mRNA transcripts were kept constant also suggests that this regulation occurs at a post-transcriptional level. PMID:24534144

  12. Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine

    PubMed Central

    Liu, Jian; Li, Chunyan; Zhang, Hua; Ma, Ping; Luo, Xianfeng; Zeng, Zhiyong; Hong, Nining; Liu, Xia; Wang, Bin; Wang, Feng; Gan, Zhenlei; Hao, Fei

    2014-01-01

    The purpose of this study was to investigate the effects of porcine interleukin (IL)-2 and IL-4 genes on enhancing the immunogenicity of a porcine reproductive and respiratory syndrome virus ORF5 DNA vaccine in piglets. Eukaryotic expression plasmids pcDNA-ORF5, pcDNA-IL-2, and pcDNA-IL-4 were constructed and then expressed in Marc-145 cells. The effects of these genes were detected using an indirect immunofluorescent assay and reverse transcription polymerase chain reaction (RT-PCR). Characteristic fluorescence was observed at different times after pcDNA-ORF5 was expressed in the Marc-145 cells, and PCR products corresponding to ORF5, IL-2, and IL-4 genes were detected at 48 h. Based on these data, healthy piglets were injected intramuscularly with different combinations of the purified plasmids: pcDNA-ORF5 alone, pcDNA-ORF5 + pcDNA-IL-2, pcDNA-ORF5 + pcDNA-IL-4, and pcDNA-ORF5 + pcDNAIL-4 + pcDNA-IL-2. The ensuing humoral immune responses, percentages of CD4+ and CD8+ T lymphocytes, proliferation indices, and interferon-? expression were analyzed. Results revealed that the piglets co-immunized with pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2 plasmids developed significantly higher antibody titers and neutralizing antibody levels, had significantly increased levels of specific T lymphocyte proliferation, elevated percentages of CD4+ and CD8+ T lymphocytes, and significantly higher IFN-? production than the other inoculated pigs (p < 0.05). PMID:24136204

  13. Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine.

    PubMed

    Tang, Deyuan; Liu, Jian; Li, Chunyan; Zhang, Hua; Ma, Ping; Luo, Xianfeng; Zeng, Zhiyong; Hong, Nining; Liu, Xia; Wang, Bin; Wang, Feng; Gan, Zhenlei; Hao, Fei

    2014-01-01

    The purpose of this study was to investigate the effects of porcine interleukin (IL)-2 and IL-4 genes on enhancing the immunogenicity of a porcine reproductive and respiratory syndrome virus ORF5 DNA vaccine in piglets. Eukaryotic expression plasmids pcDNA-ORF5, pcDNA-IL-2, and pcDNA-IL-4 were constructed and then expressed in Marc-145 cells. The effects of these genes were detected using an indirect immunofluorescent assay and reverse transcription polymerase chain reaction (RT-PCR). Characteristic fluorescence was observed at different times after pcDNA- ORF5 was expressed in the Marc-145 cells, and PCR products corresponding to ORF5, IL-2, and IL-4 genes were detected at 48 h. Based on these data, healthy piglets were injected intramuscularly with different combinations of the purified plasmids: pcDNA-ORF5 alone, pcDNA-ORF5 + pcDNA-IL-2, pcDNA-ORF5 + pcDNA-IL-4, and pcDNA-ORF5 + pcDNA- IL-4 + pcDNA-IL-2. The ensuing humoral immune responses, percentages of CD4(+) and CD8(+) T lymphocytes, proliferation indices, and interferon-g expression were analyzed. Results revealed that the piglets co-immunized with pcDNA-ORF5 + pcDNA-IL-4 + pcDNA-IL-2 plasmids developed significantly higher antibody titers and neutralizing antibody levels, had significantly increased levels of specific T lymphocyte proliferation, elevated percentages of CD4(+) and CD8(+) T lymphocytes, and significantly higher IFN-? production than the other inoculated pigs (p < 0.05). PMID:24136204

  14. Complete genome characterization of a East European Type 1 subtype 3 porcine reproductive and respiratory syndrome virus.

    PubMed

    Van Doorsselaere, J; Brar, M S; Shi, M; Karniychuk, U; Leung, F C-C; Nauwynck, H J

    2012-02-01

    Porcine reproductive and respiratory syndrome (PRRS) is a swine disease of major economic importance that causes reproductive and respiratory problems in pigs. PRRSV strains are divided into European (Type 1) and North-American (Type 2) genotypes. Within the European PRRSV genotype, three subtypes have been delineated. Full genome sequences for North American and European subtype 1 strains have been described. Here, the first complete genomic characterization of a European subtype 3 strain (Lena) is described. Amplification of Orf1a and Orf1b fragments was achieved using a set of degenerate oligonucleotides. Using RT-PCR with Lena-specific primers, the full length sequence (15001 nt) was obtained. Alignment of Lena with European subtype 1 reference strain Lelystad showed variation over the entire length (84% identity/89% similarity at amino acid level) with the most variation in Orf1a (Nsp2/NSP2) with a deletion of 29 amino acids. Phylogenetic relationships using different Orfs supported Lena's genetic distinction from European subtype 1 strains. The availability of the European subtype 3 PRRSV full genome may be important for the understanding of PRRSV evolution and the more pronounced pathogenic nature of Lena. PMID:21948018

  15. Can persistent Epstein-Barr virus infection induce Chronic Fatigue Syndrome as a Pavlov feature of the immune response?

    E-print Network

    Agliari, Elena; Vidal, Kristian Gervasi; Guerra, Francesco

    2011-01-01

    Chronic Fatigue Syndrome is a protracted illness condition (lasting even years) appearing with strong flu symptoms and with complex, systemic, defaiances by the immune system. Here we study the most widely accepted picture for its genesis, namely a persistent acute mononucleosis infection, by means of non-equilibrium statistical mechanics techniques and we show how this may drive the immune system toward an out-of-equilibrium metastable state (with long life-time) displaying chronic activation of both humoral and cellular responses: a scenario with full inflammation without a direct "causes-effect" reason. By exploiting a bridge with the neuronal scenario, we mirror killer lymphocytes $T_K$ and $B$-cells to neurons and helper lymphocytes $T_{H_1},T_{H_2}$ to synapses, hence showing that -under minimal physical assumptions- the immune system may experience the Pavlov conditional reflex phenomenon such that if the exposition to a stimulus (EBV antigens) is too long, strong internal correlations among $B,T_K,T_H...

  16. Epidemiology of Oropharyngeal Candidiasis in Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome Patients and CD4+ Counts

    PubMed Central

    Berberi, Antoine; Noujeim, Ziad; Aoun, Georges

    2015-01-01

    Background: The present study was directed to evaluate the forms of oropharyngeal candidiasis (OPC) and their correlation with CD4+ cell counts in human immunodeficiency virus (HIV) patients. Materials and Methods: This was a descriptive and analytical cross-sectional study carried out for a 2-year period, in which quantitative data collection methods were used. 50 patients with HIV infection were evaluated. Relationship between OPC and CD4+ was investigated. Results: Five different clinical forms were noticed on examination: pseudomembranous candidiasis 20/38 (P) was the most common one (52.6%) followed by erythematous 5/38 (13.15%), angular cheilitis 5/38 (13.15%) (AC), a combination of AC and E 4/38 (10.52%) or AC, E and P 4/38 (10.52%). Candida albicans was the most frequent specie isolated in 35 cases of OPC (92%). Candida tropicalis was isolated in 2 cases (5.26%) and Candida glabrata in 1 case (2.64%). The majority of patients with OPC had cell counts 28/38 (73%) <200 cells/mm3, followed by 9/38 (23%) at CD4+ cell counts of 201-499 cells/mm3. Conclusion: Oral Candida colonization and invasive infection occur more frequently in HIV-positive patient and is significantly more common in patients with CD4+ cell counts <200 cell/mm3. PMID:25878473

  17. What skills do star fund managers possess

    E-print Network

    Chen, Li-Wen

    2010-02-17

    Kosowski, Timmermann, Wermers, and White (2006) find that certain growth-oriented fund managers have substantial skill but do not stipulate the particular skills that they possess. I use novel style timing models to ...

  18. 50 CFR 648.25 - Possession restrictions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Management Measures for the Atlantic Mackerel, Squid, and Butterfish Fisheries ...Possession restrictions. (a) Atlantic mackerel. During a closure of the directed Atlantic mackerel fishery that occurs prior to June...

  19. Decriminalizing Possession of All Controlled Substances.

    ERIC Educational Resources Information Center

    Kurzman, Marc G.

    1978-01-01

    Excerpts from the Minnesota Bar Association's Blue Ribbon Committee report of findings and recommendations, with regard to dealing with possession of heroin and other controlled substances, are presented here. (Author/DS)

  20. Viruses in Antarctic lakes

    NASA Technical Reports Server (NTRS)

    Kepner, R. L. Jr; Wharton, R. A. Jr; Suttle, C. A.; Wharton RA, J. r. (Principal Investigator)

    1998-01-01

    Water samples collected from four perennially ice-covered Antarctic lakes during the austral summer of 1996-1997 contained high densities of extracellular viruses. Many of these viruses were found to be morphologically similar to double-stranded DNA viruses that are known to infect algae and protozoa. These constitute the first observations of viruses in perennially ice-covered polar lakes. The abundance of planktonic viruses and data suggesting substantial production potential (relative to bacteria] secondary and photosynthetic primary production) indicate that viral lysis may be a major factor in the regulation of microbial populations in these extreme environments. Furthermore, we suggest that Antarctic lakes may be a reservoir of previously undescribed viruses that possess novel biological and biochemical characteristics.

  1. PD-L1 expression is increased in monocyte derived dendritic cells in response to porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus infections.

    PubMed

    Richmond, O; Cecere, T E; Erdogan, E; Meng, X J; Piñeyro, P; Subramaniam, S; Todd, S M; LeRoith, T

    2015-11-15

    Host immune system suppression is thought to be crucial in the development of porcine circovirus associated diseases (PCVAD). Many immune suppressive mechanisms have been studied in cases of PCVAD, however, the role of programmed death ligand-1 (PD-L1) during porcine circovirus type 2 (PCV2) infection and PCVAD development has yet to be determined. PD-L1 has become an important research target because of its ability to interfere with effective T-cell activity and proliferation during the course of an immune response. In this study, porcine monocyte derived dendritic cells (MoDC) were infected with different combinations of PCV2 and porcine reproductive and respiratory syndrome virus (PRRSV) and evaluated for expression levels of PD-L1, as well as the expression levels of swine major histocompatibility complexes 1 and 2 (SLA-1 and SLA-2) as a measure of MoDC stimulatory capacity. PD-L1 expression levels were also tested in MoDCs after treatment with interferon alpha (IFN-?) and beta (IFN-?). The results showed that the expression levels of PD-L1 were increased in PCV2-infected MoDCs, as well as in PCV2 and PRRSV co-infected MoDCs. The MoDCs infected with PRRSV only also showed a strain-dependent increase in PD-L1 expression. Both IFN-? and IFN-? treatment also increased the expression levels of PD-L1 in MoDCs. SLA-1 and 2 expression levels were increased by PCV2 infection, and altered in the PRRSV, and PCV2+PRRSV co-infected MoDCs in a strain-dependent manner. These results indicate a potential immuno-suppressive role for dendritic cells during PCV2 infection and the development of PCVAD and will be helpful in more fully elucidating the underlying mechanisms leading to clinical PCVAD. PMID:26553563

  2. Immune responses of whiteleg shrimp, Litopenaeus vannamei (Boone, 1931), to bacterially expressed dsRNA specific to VP28 gene of white spot syndrome virus.

    PubMed

    Taju, G; Madan, N; Abdul Majeed, S; Kumar, T Raj; Thamizhvanan, S; Otta, S K; Sahul Hameed, A S

    2015-05-01

    In this study, dsRNA specific to VP28 gene of white spot syndrome virus (WSSV) of shrimp was synthesized in Escherichia coli in large scale and studied the immune response of shrimp to dsRNA-VP28. The haematological parameters such as clotting time and total haemocytes counts, and immunological parameters such as prophenoloxidase (proPO), superoxide dismutase (SOD), superoxide anion (SOA) and malondialdehyde content, as well as the mRNA expression of ten immune-related genes were examined to estimate the effect of dsRNA-VP28 on the innate immunity of Litopenaeus vannamei. The activities of proPO, SOA and SOD significantly increased in haemocyte after dsRNA-VP28 treatment, whereas MDA content did not change significantly. Among the ten immune-related genes examined, only the mRNA expression of proPO, cMnSOD, haemocyanin, crustin, BGBP, lipopolysaccharides (LPs), lectin and lysozyme in haemocytes, gill and hepatopancreas of L. vannamei, was significantly upregulated at 12 h after dsRNA-VP28 treatment, while no significant expression changes were observed in Toll receptor and tumour receptor genes. The increase of proPO and SOD activities, and SOA level and mRNA expression level of proPO, cMnSOD, haemocyanin, crustin, BGBP, LPs, lectin and lysozyme after dsRNA-VP28 stimulation indicate that these immune-related genes were involved in dsRNA-VP28-induced innate immunity in shrimp. PMID:24917208

  3. Comparative Effects of Vaccination against Porcine Circovirus Type 2 (PCV2) and Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) in a PCV2-PRRSV Challenge Model

    PubMed Central

    Park, Changhoon; Oh, Yeonsu; Seo, Hwi Won; Han, Kiwon

    2013-01-01

    The objective of the present study was to determine the effects of porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) vaccinations in an experimental PCV2-PRRSV challenge model, based on virological (viremia), immunological (neutralizing antibodies [NAs], gamma interferon-secreting cells [IFN-?-SCs], and CD4+ CD8+ double-positive cells), and pathological (lesions and antigens in lymph nodes and lungs) evaluations. A total of 72 pigs were randomly divided into 9 groups (8 pigs per group): 5 vaccinated and challenged groups, 3 nonvaccinated and challenged groups, and a negative-control group. Vaccination against PCV2 induced immunological responses (NAs and PCV2-specific IFN-?-SCs) and reduced PCV2 viremia, PCV2-induced lesions, and PCV2 antigens in the dually infected pigs. However, vaccination against PCV2 did not affect the PRRSV immunological responses (NAs and PRRSV-specific IFN-?-SCs), PRRSV viremia, PRRSV-induced lesions, or PRRSV antigens in the dually infected pigs. Vaccination against PRRSV did not induce immunological responses (PRRSV-specific IFN-?-SCs) or reduce PRRSV viremia, PRRSV-induced lesions, or PRRSV antigens in the dually infected pigs. In addition, vaccination against PRRSV increased PCV2 viremia, PCV2-induced lesions, and PCV2 antigens in the dually infected pigs. In summary, vaccination against PCV2 reduced PCV2 viremia, PCV2-induced lesions, and PCV2 antigens in the dually infected pigs. However, vaccination against PRRSV increased PCV2 viremia, PCV2-induced lesions, and PCV2 antigens in the dually infected pigs. Therefore, the PCV2 vaccine decreased the potentiation of PCV2-induced lesions by PRRSV in dually infected pigs. In contrast, the PRRSV vaccine alone did not decrease the potentiation of PCV2-induced lesions by PRRSV in dually infected pigs. PMID:23302743

  4. The shrimp IKK–NF-?B signaling pathway regulates antimicrobial peptide expression and may be subverted by white spot syndrome virus to facilitate viral gene expression

    PubMed Central

    Wang, Pei-Hui; Gu, Zhi-Hua; Wan, Ding-Hui; Liu, Bo-Du; Huang, Xian-De; Weng, Shao-Ping; Yu, Xiao-Qiang; He, Jian-Guo

    2013-01-01

    The I?B kinases IKK? and IKK? and the IKK-related kinases TANK-binding kinase 1 (TBK1) and IKK? are the master regulators of the NF-?B signaling pathway. Although this pathway has been extensively studied in mammals, less attention has been paid in crustaceans, which have significant economic value. Here, we report the cloning and functional studies of two IKK homologs, LvIKK? and LvIKK?, from Pacific white shrimp, Litopenaeus vannamei. LvIKK? and LvIKK? mRNAs are widely expressed in different tissues and are responsive to white spot syndrome virus (WSSV) infection. When overexpressed in Drosophila S2 cells, LvIKK? but not LvIKK? activates the promoters of NF-?B pathway-controlled antimicrobial peptide genes (AMPs), such as the Penaeidins (PENs). In HEK 293T cells, both LvIKK? and LvIKK? activate an NF-?B reporter. The silencing of LvIKK? or LvIKK? using double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) decreases the expression of L. vannamei AMPs, including PENs, lysozyme and crustins. Intriguingly, LvIKK?- or LvIKK?-silenced L. vannamei are resistant to WSSV infection. We hypothesized that successful infection with WSSV requires the activation of the IKK–NF-?B signaling pathway to modulate viral gene expression. We constructed luciferase reporters for 147 WSSV genes. By screening, we found that the WSV051, WSV059, WSV069, WSV083, WSV090, WSV107, WSV244, WSV303, WSV371 and WSV445 promoters can be activated by LvIKK? or LvIKK? in Drosophila S2 cells. Taken together, our results reveal that LvIKK? and LvIKK? may participate in the regulation of shrimp AMPs and that WSSV may subvert the L. vannamei IKK–NF-?B signaling pathway to facilitate viral gene expression. PMID:23954949

  5. Cloning and characterization of a caspase gene from black tiger shrimp (Penaeus monodon)-infected with white spot syndrome virus (WSSV).

    PubMed

    Wongprasert, Kanokpan; Sangsuriya, Pakkakul; Phongdara, Amornrat; Senapin, Saengchan

    2007-08-01

    A black tiger shrimp (Penaeus monodon) caspase cDNA homologue (PmCasp) has been identified from a hemocyte library using a previously identified caspase homologue from the banana shrimp (Penaeus merguiensis) as a probe. The full-length PmCasp was 1202bp with a 954bp open reading frame, encoding 317 amino acids. The deduced protein contained a potential active site (QACRG pentapeptide) conserved in most caspases. It had 83% identity with caspase of P. merguiensis and 30% identity with drICE protein of Drosophila melanogaster, and it exhibited caspase-3 activity in vitro. PmCasp was cloned and expressed in Escherichia coli and a rabbit polyclonal antiserum was produced. In Western blots, the antiserum reacted with purified recombinant PmCasp and with lysates of E. coli containing the expressed plasmid. In crude protein extracts from normal shrimp, the antiserum reacted with 36 and 26kDa bands likely to correspond to inactive pro-caspase and its proteolytic intermediate form, respectively. PmCasp expression was measured in normal shrimp and in white spot syndrome virus (WSSV)-infected shrimp at 24 and 48h post-injection (p.i.) by semi-quantitative RT-PCR, Western blot analysis, and immunohistochemistry. Semi-quantitative RT-PCR analysis revealed up-regulation of PmCasp at 48h p.i. and expression remained high up to the moribund state. These results were supported by Western blot analysis showing increased PmCasp protein levels at 24 and 48h p.i. when compared to normal control shrimp. Immunohistochemical analysis of gills from the WSSV-infected shrimp revealed immunoreactivity localized in the cytoplasm of both normal and apparently apoptotic cells. In summary, a caspase-3 like gene is conserved in P. monodon and is up-regulated after WSSV infection. PMID:17617486

  6. Detection of Porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in oral fluid specimens using a commercial PRRSV serum antibody enzyme-linked immunosorbent assay.

    PubMed

    Kittawornrat, Apisit; Prickett, John; Wang, Chong; Olsen, Chris; Irwin, Christa; Panyasing, Yaowalak; Ballagi, Andrea; Rice, Anna; Main, Rodger; Johnson, John; Rademacher, Chris; Hoogland, Marlin; Rowland, Raymond; Zimmerman, Jeffrey

    2012-03-01

    The purpose of the present study was to evaluate the diagnostic performance of a commercial serum antibody enzyme-linked immunosorbent assay (ELISA) modified to detect anti-Porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in pen-based oral fluid specimens. Experimental and field oral fluid samples of defined status in reference to exposure of swine with PRRSV were used to derive the kinetics of detectable concentrations of antibody against PRRSV. Immunoglobulin (Ig)M and IgA were readily detected in oral fluid specimens from populations in which PRRSV infection was synchronized among all individuals but not in samples collected in commecial herds. In contrast, IgG was readily detected at diagnostically useful levels in both experimental and field samples for up to 126 days. Estimates of the IgG oral fluid ELISA performance were based on results from testing positive oral fluid samples (n = 492) from experimentally inoculated pigs (n = 251) and field samples (n = 241) and negative oral fluid samples (n = 367) from experimentally inoculated pigs (n = 84) and field samples (n = 283). Receiver operating characteristic analysis estimated the diagnostic sensitivity and specificity of the assay as 94.7% (95% confidence interval [CI]: 92.4, 96.5) and 100% (95% CI: 99.0, 100.0), respectively, at a sample-to-positive ratio cutoff of ?0.40. The results of the study suggest that the IgG oral fluid ELISA can provide efficient, cost-effective PRRSV monitoring in commercial herds and PRRSV surveillance in elimination programs. PMID:22379043

  7. Comparison of commercial enzyme-linked immunosorbent assays and fluorescent microbead immunoassays for detection of antibodies against porcine reproductive and respiratory syndrome virus in boars.

    PubMed

    Gerber, Priscilla F; Giménez-Lirola, Luis G; Halbur, Patrick G; Zhou, Lei; Meng, Xiang-Jin; Opriessnig, Tanja

    2014-03-01

    The objective of this study was to compare the ability of two commercial enzyme-linked immunosorbent assays (ELISAs) and an in-house fluorescent microbead immunoassay (FMIA) to detect IgG antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) types 1 and 2 in serum and oral fluids from boars infected experimentally. Samples from uninfected control pigs and PRRSV-negative field samples were also used. Serum samples were tested by ELISAs (IDEXX Se, HIPRA Se) and an in-house FMIA-Se for detection of PRRSV types 1 and 2. Oral fluids were tested by ELISAs (IDEXX-SO, IDEXX-OF, HIPRA-OF) for detection of PRRSV types 1 and 2. Among the sera, IDEXX-Se and HIPRA-Se had similar sensitivity and specificity (p>0.05); however, IDEXX-Se detected positive animals earlier than HIPRA-Se (p<0.05). FMIA-Se had the highest false-positive rates in known negative field samples (1/205 for IDEXX-Se, 5/205 for HIPRA-Se, and 37/205 for FMIA-Se; p<0.01). Serum and oral fluid samples had similar detection rates and antibody kinetics using the IDEXX tests. There was a higher detection rate in serum than oral fluid using the HIPRA assays. In this study, the nucleocapsid protein utilized as antigen in the FMIAs yielded a low specificity. IDEXX-Se had the earliest detection and similar sensitivity and specificity to the HIPRA-Se. PMID:24361873

  8. Ring test evaluation of the repeatability and reproducibility of a Porcine reproductive and respiratory syndrome virus oral fluid antibody enzyme-linked immunosorbent assay.

    PubMed

    Kittawornrat, Apisit; Wang, Chong; Anderson, Gary; Ballagi, Andrea; Broes, André; Carman, Suzanne; Doolittle, Kent; Galeota, Judith; Johnson, John; Lizano, Sergio; Nelson, Eric; Patnayak, Devi; Pogranichniy, Roman; Rice, Anna; Scherba, Gail; Zimmerman, Jeffrey

    2012-11-01

    The precision of a Porcine reproductive and respiratory syndrome virus (PRRSV) oral fluid antibody enzyme-linked immunosorbent assay (ELISA) was evaluated by calculating reliability coefficients for assay repeatability (within laboratory) and assay reproducibility (between laboratories). Randomly ordered oral fluid samples of known (n = 39) and unknown (n = 224) PRRSV antibody status were tested in 12 diagnostic laboratories. Each laboratory tested the samples twice, first using an antibody ELISA kit and reagents provided to them (phase 1) and then using an ELISA kit and reagents configured in their respective laboratory (phase 2). Repeatability (within laboratory) reliability coefficients calculated using results from samples of known PRRSV antibody status ranged from 0.724 to 0.997 in phase 1 and from 0.953 to 0.998 in phase 2. Reproducibility (between laboratories) reliability coefficients were calculated for 3 conditions: case 1--samples of unknown status (n = 224); case 2--samples of known status (n = 39), and case 3--all samples (n = 263). Among the 3 cases, reliability coefficients ranged from 0.937 to 0.964 in phase 1 and from 0.922 to 0.935 in phase 2. For case 3, it was estimated that 96.67% of the total variation in phase 1 and 93.21% in phase 2 could be attributed to the oral fluid samples themselves. Overall, the PRRSV oral fluid antibody ELISA was highly repeatable and reproducible. The current study supports the routine use of this test in laboratories providing diagnostic service to pig producers. PMID:22964428

  9. The novel white spot syndrome virus-induced gene, PmERP15, encodes an ER stress-responsive protein in black tiger shrimp, Penaeus monodon.

    PubMed

    Leu, Jiann-Horng; Liu, Kuan-Fu; Chen, Kuan-Yu; Chen, Shu-Hwa; Wang, Yu-Bin; Lin, Chung-Yen; Lo, Chu-Fang

    2015-04-01

    By microarray screening, we identified a white spot syndrome virus (WSSV)-strongly induced novel gene in gills of Penaeus monodon. The gene, PmERP15, encodes a putative transmembrane protein of 15?kDa, which only showed some degree of similarity (54-59%) to several unknown insect proteins, but had no hits to shrimp proteins. RT-PCR showed that PmERP15 was highly expressed in the hemocytes, heart and lymphoid organs, and that WSSV-induced strong expression of PmERP15 was evident in all tissues examined. Western blot analysis likewise showed that WSSV strongly up-regulated PmERP15 protein levels. In WSSV-infected hemocytes, immunofluorescence staining showed that PmERP15 protein was colocalized with an ER enzyme, protein disulfide isomerase, and in Sf9 insect cells, PmERP15-EGFP fusion protein colocalized with ER -Tracker™ Red dye as well. GRP78, an ER stress marker, was found to be up-regulated in WSSV-infected P.?monodon, and both PmERP15 and GRP78 were up-regulated in shrimp injected with ER stress inducers tunicamycin and dithiothreitol. Silencing experiments showed that although PmERP15 dsRNA-injected shrimp succumbed to WSSV infection more rapidly, the WSSV copy number had no significant changes. These results suggest that PmERP15 is an ER stress-induced, ER resident protein, and its induction in WSSV-infected shrimp is caused by the ER stress triggered by WSSV infection. Furthermore, although PmERP15 has no role in WSSV multiplication, its presence is essential for the survival of WSSV-infected shrimp. PMID:25499032

  10. Bayesian analysis of risk factors for infection with a genotype of porcine reproductive and respiratory syndrome virus in Ontario swine herds using monitoring data.

    PubMed

    Kwong, Grace P S; Poljak, Zvonimir; Deardon, Rob; Dewey, Cate E

    2013-07-01

    Porcine reproductive and respiratory syndrome (PRRS) has a worldwide distribution. This economically important endemic disease causes reproductive failure in breeding stock and respiratory tract illness in young pigs. In Ontario restricted fragment length polymorphism (RFLP) 1-18-4 has been determined as one of the most common virus genotypes. Individual-level models (ILMs) for infectious diseases, fitted in a Bayesian MCMC framework, have been used to describe both the spatial and temporal spread of diseases. They are an intuitive and flexible class of models that can take into account population heterogeneity via various individual-level covariates. The objective of this study is to identify relative importance of risk factors for the spread of the genotype 1-18-4 from monitoring data in southern Ontario using ILMs. Specifically, we explore networks through which resources are obtained or delivered, as well as the ownership structure of herds, and identify factors that may be contributing to high risk of infection. A population of 316 herds which experienced their PRRS outbreaks between September 2004 and August 2007 are included in the analyses, in which 194 (61%) are sow herds. During the study period, 45 herds (27 sow herds) experienced their first outbreak due to RFLP 1-18-4. Our results show that the three relatively most important factors for the spread of 1-18-4 genotype in Ontario swine herds were sharing the same herd ownership, gilt source and market trucks. All other networks had relatively smaller impact on spread of this PRRSV genotype. Spatial proximity could not be identified as important contributor to spread. Our findings also suggest that gilt acclimation should be practiced whenever possible and appropriate to reduce the risk for the herd and for others as it is already widely implemented and recommended in the North American swine industry. PMID:23416041

  11. Influence of the home environment on the prevention of mother to child transmission of human immunodeficiency virus/acquired immune-deficiency syndrome in South Africa.

    PubMed

    Sewnunan, A; Modiba, L M

    2015-12-01

    The human immunodeficiency virus and acquired immune-deficiency syndrome (HIV/AIDS) is still a 'family crises' which marks the beginning of the deterioration of the family unit and the trauma in the emotional, psychological and material lives of both the mother and child. In South African context where the majority of HIV-positive mothers are young single women who live in extended families, disclosure to the sexual partner alone is not an adequate condition for the success of prevention of mother to child transmission (PMTCT). In South Africa, close to one in three women who attend antenatal clinics are HIV positive. KwaZulu-Natal is one of the worst affected provinces, where as many as 40-60% of pregnant women attending antenatal services are living with HIV infection. The study sought to investigate the link between the home environment and its contribution to the success of the programme on PMTCT of HIV/AIDS. A qualitative, explorative, descriptive and contextual study was used in this study to explore whether the home environment for the support system is available for the HIV-positive women on the PMTCT programme. The population of this study included all women who have undergone counselling and tested HIV positive and who have joined the programme on PMTCT of HIV/AIDS in a specific hospital in KwaZulu-Natal Province. Although 14 women agreed to participate in the study, only 10 women were interviewed as saturation was attained. Data were collected using semi-structured interview schedule. Interviews were audio-taped and field notes were taken. Content analysis was used and it was done manually. This study revealed that one of the major issues still surrounding HIV/AIDS and PMTCT is that of non-disclosure, selective disclosure and the stigma and discrimination that surrounds this disease. PMID:26694631

  12. Histoplasmosis in patients with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS): multicenter study of outcomes and factors associated with relapse.

    PubMed

    Myint, Thein; Anderson, Albert M; Sanchez, Alejandro; Farabi, Alireza; Hage, Chadi; Baddley, John W; Jhaveri, Malhar; Greenberg, Richard N; Bamberger, David M; Rodgers, Mark; Crawford, Timothy N; Wheat, L Joseph

    2014-01-01

    Although discontinuation of suppressive antifungal therapy for acquired immunodeficiency syndrome (AIDS)-associated histoplasmosis is accepted for patients with immunologic recovery, there have been no published studies of this approach in clinical practice, and minimal characterization of individuals who relapse with this disease. We performed a multicenter retrospective cohort study to determine the outcome in AIDS patients following discontinuation of suppressive antifungal therapy for histoplasmosis. Ninety-seven patients were divided into a physician-discontinued suppressive therapy group (PD) (38 patients) and a physician-continued suppressive therapy group (PC) (59 patients). The 2 groups were not statistically different at baseline, but at discontinuation of therapy and at the most recent follow-up there were significant differences in adherence to therapy, human immunodeficiency virus (HIV) RNA, and urinary Histoplasma antigen concentration. There was no relapse or death attributed to histoplasmosis in the PD group compared with 36% relapse (p < 0.0001) and 5% death (p = 0.28) in the PC group. Relapse occurred in 53% of the nonadherent patients but not in the adherent patients (p < 0.0001). Sixty-seven percent of patients with initial central nervous system (CNS) histoplasmosis relapsed compared to 15% of patients without CNS involvement (p = 0.0004), which may be accounted for by nonadherence. In addition, patients with antigenuria above 2.0 ng/mL at 1-year follow-up were 12.82 times (95% confidence interval, 2.91-55.56) more likely to relapse compared to those with antigenuria below 2.0 ng/mL. Discontinuation of antifungal therapy was safe in adherent patients who completed at least 1 year of antifungal treatment, and had CD4 counts >150 cells/mL, HIV RNA <400 c/mL, Histoplasma antigenuria <2 ng/mL (equivalent to <4.0 units in second-generation method), and no CNS histoplasmosis. PMID:24378739

  13. Genetic Susceptibility Is One of the Determinants for Severe Fever with Thrombocytopenia Syndrome Virus Infection and Fatal Outcome: An Epidemiological Investigation.

    PubMed

    Sun, Jimin; Tang, Yuming; Ling, Feng; Chang, Yue; Ye, Xiaohong; Shi, Wen; Zhang, Lei; Chen, Zhiping; Lin, Haijiang; Qiu, Zaiping; Zhang, Yanjun; Zhang, Rong; Mao, Haiyan; Chen, Enfu; Lin, Junfen; Jiang, Jianmin; Xia, Shichang; Gong, Zhenyu

    2015-01-01

    Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease in China and case-fatality rate of SFTS is very high (approximately 10%). However, genetic susceptibility for SFTS virus (SFTSV) infection and fatal outcome of SFTSV infection in humans are unclear. In this study, we investigated the clinical, laboratory and epidemiological features of SFTS in a cluster of three sisters who died of SFTSV infection between late April and mid-May 2014. Before disease onset, two of the sisters (Case A and case B) had common exposure history for ticks by working together in a field to pick tea leaves from April 8 to April 12. The third sister (Case C) did not live or work together with case A and B, but had ticks in her living environment. SFTSV RNA sequences were amplified from three cases were not identical, suggesting that the three sisters were most likely infected with SFTSV through tick bite rather than through person-to-person transmission of SFTSV. The sequence of SFTSV from case C was identical to SFTSV sequences from 3 groups of ticks collected around the residential area of case C. Seroprevalence of SFTSV IgG antibody among healthy population in the area where the patients resided was 4.05% (3/74). The majority of SFTSV infections were mild cases and all three sisters died of SFTSV infection suggested that they were highly susceptible to SFTSV. Our findings indicated that genetic susceptibility was a risk factor for SFTSV infection and fatal outcome. PMID:26207638

  14. Genetic Susceptibility Is One of the Determinants for Severe Fever with Thrombocytopenia Syndrome Virus Infection and Fatal Outcome: An Epidemiological Investigation

    PubMed Central

    Chang, Yue; Ye, Xiaohong; Shi, Wen; Zhang, Lei; Chen, Zhiping; Lin, Haijiang; Qiu, Zaiping; Zhang, Yanjun; Zhang, Rong; Mao, Haiyan; Chen, Enfu; Lin, Junfen; Jiang, Jianmin; Xia, Shichang; Gong, Zhenyu

    2015-01-01

    Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease in China and case-fatality rate of SFTS is very high (approximately 10%). However, genetic susceptibility for SFTS virus (SFTSV) infection and fatal outcome of SFTSV infection in humans are unclear. In this study, we investigated the clinical, laboratory and epidemiological features of SFTS in a cluster of three sisters who died of SFTSV infection between late April and mid-May 2014. Before disease onset, two of the sisters (Case A and case B) had common exposure history for ticks by working together in a field to pick tea leaves from April 8 to April 12. The third sister (Case C) did not live or work together with case A and B, but had ticks in her living environment. SFTSV RNA sequences were amplified from three cases were not identical, suggesting that the three sisters were most likely infected with SFTSV through tick bite rather than through person-to-person transmission of SFTSV. The sequence of SFTSV from case C was identical to SFTSV sequences from 3 groups of ticks collected around the residential area of case C. Seroprevalence of SFTSV IgG antibody among healthy population in the area where the patients resided was 4.05% (3/74). The majority of SFTSV infections were mild cases and all three sisters died of SFTSV infection suggested that they were highly susceptible to SFTSV. Our findings indicated that genetic susceptibility was a risk factor for SFTSV infection and fatal outcome. PMID:26207638

  15. Nonstructural protein 1{alpha} subunit-based inhibition of NF-{kappa}B activation and suppression of interferon-{beta} production by porcine reproductive and respiratory syndrome virus

    SciTech Connect

    Song Cheng; Krell, Peter; Yoo, Dongwan

    2010-11-25

    Induction of type I interferon (IFN-{alpha}/{beta}) is an early antiviral response of the host, and porcine reproductive and respiratory syndrome virus (PRRSV) has been reported to downregulate the IFN response during infection in cells and pigs. We report that the PRRSV nonstructural protein 1{alpha} (Nsp1{alpha}) subunit of Nsp1 is a nuclear-cytoplasmic protein distributed to the nucleus and contains a strong suppressive activity for IFN-{beta} production that is mediated through the retinoic acid-inducible gene I (RIG-I) signaling pathway. Nsp1{alpha} suppressed the activation of nuclear factor (NF)-{kappa}B when stimulated with dsRNA or tumor necrosis factor (TNF)-{alpha}, and NF-{kappa}B suppression was RIG-I-dependent. The suppression of NF-{kappa}B activation was associated with the poor production of IFN-{beta} during PRRSV infection. The C-terminal 14 amino acids of the Nsp1{alpha} subunit were critical in maintaining immunosuppressive activity of Nsp1{alpha} for both IFN-{beta} and NF-{kappa}B, suggesting that the newly identified zinc finger configuration comprising of Met180 may be crucial for inhibitory activities. Nsp1{alpha} inhibited I{kappa}B phosphorylation and as a consequence NF-{kappa}B translocation to the nucleus was blocked, leading to the inhibition of NF-{kappa}B stimulated gene expression. Our results suggest that PRRSV Nsp1{alpha} is a multifunctional nuclear protein participating in the modulation of the host IFN system.

  16. Comparison of two genetically distant type 2 porcine reproductive and respiratory syndrome virus (PRRSV) modified live vaccines against Vietnamese highly pathogenic PRRSV.

    PubMed

    Do, Duy Tien; Park, Changhoon; Choi, Kyuhyung; Jeong, Jiwoon; Nguyen, Toan Tat; Nguyen, Khang Duong; Vo, Dai Tan; Chae, Chanhee

    2015-09-30

    Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) known as pig high fever disease was first reported in China and has spread rapidly in neighboring southeastern Asian countries. The objective of this study was to evaluate the efficacy of a new type 2 PRRSV modified live vaccine (vaccine A) against a challenge with a HP-PRRSV and to compare the efficacy of two genetically distant type 2 PRRSV modified vaccines (vaccine A for lineage 8 and vaccine B for lineage 5) against HP-PRRSV (lineage 8) challenge. Pigs were divided into 4 groups (n=12/group); vaccinated challenged (2 groups), unvaccinated challenged, and unvaccinated unchallenged groups. Regardless of vaccines, vaccinated challenged pigs showed significantly lower (P<0.05) mean rectal temperatures and respiratory scores, levels of HP-PRRSV viremia, and lung lesions and HP-PRRSV antigens within lung lesions compared to unvaccinated challenged pigs. Vaccinated challenged pigs had significantly higher (P<0.05) numbers of interferon-? secreting cells (IFN-?-SC) compared to unvaccinated challenged pigs. Significant differences were also found when comparing two type 2 PRRSV vaccines after HP-PRRSV challenge. The use of type 2 PRRSV vaccine A was able to significantly reduce fever when compared to type 2 PRRSV vaccine B in vaccinated challenged pigs. Vaccination of pigs with vaccine A reduced viral loads in their blood and induced higher numbers of HP-PRRSV-specific IFN-?-SC than vaccination of pigs with vaccine B. This study demonstrates partial protection of two genetically distant type 2 PRRSV vaccines against HP-PRRSV challenge in growing pigs. PMID:26149103

  17. [The development of a rapid loop-mediated indirect PCR method for detection and differentiation of highly and lowly pathogenic porcine reproductive and respiratory syndrome virus].

    PubMed

    Zheng, Ming; Li, Hua-Wei; Bian, Chuan-Zhou; Wang, Yong-Fen; Wang, Lao-Qi

    2013-06-01

    The aim of this study is to establish the method of loop-mediated indirect PCR assay for detection of Reproductive and Respiratory Syndrome Virus (PRRSV) infection and differentiation of highly pathogenic PRRSV (HP-PRRSV) and lowly pathogenic PRRSV (LP-PRRSV). Based on the alignments of ORF2 gene sequences and ORFla gene sequences of PRRSV Chinese isolates deposited in GenBank, two pairs of specific probes were designed and labeled to both ends of the soybean Lectin gene fragment by PCR, respectively. The probe-labeled soybean Lectin genes were used to be reporter genes for detection and differentiation of PRRSV. After one round strand displacement reaction, the reporter genes were amplified by reverse PCR. The specific PCR products were 193bp, 355bp for HP-PRRSV and 193bp, 442bp for LP-PRRSV, respectively. The method could detect 5. 6 TCID50/mL LP-PRRSV RNA and 18 TCIDs0/ mL HP-PRRSV RNA, and co-infection did not affect detection sensitivity. No amplification was observed with other porcine originated pathogens including CSFV, PPV, PRV, PCV2, ETEC and Haemophilus parasui. Twenty clinical samples were used for comparative testing with conventional PCR. Fourteen samples were found positive for PRRSV by the loop-mediated indirect PCR, of which 4 were LP-PRRSV, 9 HP-PRRSV and 1 LP/HP-PRRSV co-infection, consistent with the conventional PCR test results. In conclusion, the loop-mediated indirect PCR is a simple, rapid, sensitive and specific etiologic diagnosis tool, and suitable for the differential diagnosis of HP/LP-PRRSV, especially for identification of mixed infection of HP/LP-PRRSV. PMID:23894998

  18. Comparison of RNA extraction and real-time reverse transcription polymerase chain reaction methods for the detection of Porcine reproductive and respiratory syndrome virus in porcine oral fluid specimens.

    PubMed

    Chittick, Wayne A; Stensland, Wendy R; Prickett, John R; Strait, Erin L; Harmon, Karen; Yoon, Kyoung-Jin; Wang, Chong; Zimmerman, Jeffrey J

    2011-03-01

    The objective of the current study was to evaluate various RNA extraction and polymerase chain reaction (PCR) protocols for the detection of Porcine reproductive and respiratory syndrome virus (PRRSV) in porcine oral fluids. Extraction protocols were selected based on ease of use and compatibility with high-throughput, automated systems. The results showed marked differences among extraction protocols, PCR protocols, and combinations thereof in detecting PRRSV in the oral fluid matrix. An important finding was that PCR reactions were partially inhibited by unknown factors in the oral fluid matrix and that inhibition was reduced by use of a higher concentration of PCR enzymes. The results suggest that further optimization of PCR assays for porcine oral fluids is needed and that laboratories should not assume that methods optimized for detection of virus in serum will perform equally with porcine oral fluids. PMID:21398443

  19. Spatial and temporal patterns of porcine reproductive and respiratory syndrome virus (PRRSV) genotypes in Ontario, Canada, 2004–2007

    PubMed Central

    2014-01-01

    Background The spread of PRRSV among pig herds has been investigated experimentally, but few observational studies have investigated this subject. Because PRRSV is endemic and live modified vaccines are used in Ontario, the spatial and temporal distributions of 6 PRRSV genotypes were investigated in the province during the period from 2004–2007. The purpose was to find evidence of spread of PRRSV genotypes and determine if spread could be attributed to supplier or ownership connections between herds. Sequence information from PRRSV ORF5 and related source-herd demographic information were obtained from diagnostic submissions to the Animal Health Laboratory, University of Guelph. Results A spatial cluster that could not be attributed to supplier or ownership connections among herds in the cluster was detected for RFLP type 1-3-4. Because of genetic dissimilarity among members of the cluster, it was considered to be a result of past spread of the RFLP type. A spatio-temporal cluster detected for RFLP type 1-18-4 was attributed to a shared gilt supplier among the herds in the cluster. Significant spatio-temporal patterns detected for RFLP type 2-5-2, which is considered to be a vaccine-type virus were most likely due to grouping of herds in an ownership that used the corresponding vaccine. Clustering within herd-ownership was a risk factor for presence of five of the six genotypes investigated in the present study. Conclusions Although the literature indicates that PRRSV can spread via aerosol between pig herds, the present study found no strong evidence of this occurring in Ontario. The evidence pointed toward transmission of PRRSV occurring in this population by common sources of animals or similarity of herd ownership, which is a proxy measure for other connections between herds. It is also apparent that the recognition and testing of these connections between herds is a necessary part of interpreting spatio-temporal patterns of PRRSV genotypes. PMID:24708804

  20. [Prevalence of antibodies against the viruses of European swine fever, Aujeszky's disease and "porcine reproductive and respiratory syndrome" in wild boars in the federal states Sachsen-Anhalt and Brandenburg].

    PubMed

    Oslage, U; Dahle, J; Müller, T; Kramer, M; Beier, D; Liess, B

    1994-01-01

    During the hunting season from 1991/1992 blood samples were collected from wild boar shot in the Federal States of Sachsen-Anhalt (482 samples) and Brandenburg (177 samples) which corresponds to 2.1 and 0.4% of the total hunting bag. All sera were screened in a complex trapping blocking (CTB) ELISA for antibodies against hog cholera virus (HCV) and in an indirect ELISA for antibodies against Aujeszky's disease virus (ADV). Additionally the sera were tested for neutralizing antibodies against HCV strain ALFORT/187, bovine viral diarrhoea virus (BVDV) strains NADL and 1138/69, and against an ADV field isolate. In case of questionable results sera were tested against HCV strain "BERGEN", HCV vaccine strain "RIEMS" and three HCV field isolates from wild boar. The serological testing for antibodies against "porcine reproductive and respiratory syndrome virus" (PRRSV) was carried out in indirect immunoperoxidase monolayer assay (IPMA). Four sera (Sachsen-Anhalt) reacted positive in CTB-ELISA. Seven sera yielded neutralizing antibodies against HCV but only one of the "non-negative" samples scored positive in both techniques, ELISA and VNT. Two sera (Brandenburg) had low neutralizing antibody titres against Alfort/197 but scored negative in CTB-ELISA. Screening for antibodies against ADV of 640 sera led to 13 positive sera including 5 positive findings in both ELISA and VNT. Antibodies against PRRSV were detected in two sera which were collected Sachsen-Anhalt. Estimations resulted in a prevalence of about 5% for antibodies against HCV. PMID:8131731

  1. Original Contribution Confidence Intervals for Biomarker-based Human Immunodeficiency Virus

    E-print Network

    Cole, Stephen R.

    , acquired immunodeficiency syndrome; CI, confidence interval; HIV, human immunodeficiency virus; STARHSOriginal Contribution Confidence Intervals for Biomarker-based Human Immunodeficiency Virus biologic specimens can be used to estimate human immunodeficiency virus (HIV) incidence using a two

  2. Possession divestment by sales in later life.

    PubMed

    Ekerdt, David J; Addington, Aislinn

    2015-08-01

    Residential relocation in later life is almost always a downsizing, with many possessions to be divested in a short period of time. This article examines older movers' capacities for selling things, and ways that selling attenuates people's ties to those things, thus accomplishing the human dis-possession of the material convoy. In qualitative interviews in 79 households in the Midwestern United States, older adults reported their experience with possession sales associated with residential relocation. Among this group, three-quarters of the households downsized by selling some belongings. Informal sales seemed the least fraught of all strategies, estate sales had mixed reviews, and garage sales were recalled as laborious. Sellers' efforts were eased by social relations and social networks as helpers and buyers came forward. As selling proceeded, sentiment about possessions waned as their materiality and economic value came to the fore, easing their detachment from the household. Possession selling is challenging because older adults are limited in the knowledge, skills, and efforts that they can apply to the recommodification of their belongings. Selling can nonetheless be encouraged as a divestment strategy as long as the frustrations and drawbacks are transparent, and the goal of ridding is kept in view. PMID:26162722

  3. Additive inhibition of porcine reproductive and respiratory syndrome virus infection with the soluble sialoadhesin and CD163 receptors.

    PubMed

    Chen, Yang; Guo, Rui; He, Shan; Zhang, Xinyu; Xia, Xiaoli; Sun, Huaichang

    2014-01-22

    Porcine reproductive and respiratory syndrome (PRRS) is an economically important swine disease to the swine industry worldwide. Current PRRS vaccines are only partially effective and new vaccine development faces great challenges. Sialoadhesin (Sn) and CD163 are the two essential receptors for PRRSV infection of porcine alveolar macrophage (PAM). To investigate the feasibility of the soluble viral receptors for PRRS control, in the present study we generated recombinant adenovirus (rAd) expressing the four N-terminal Ig-like domains of porcine Sn (Sn4D), the fifth SRCR domain (SRCR5) or domains 5-9 (SRCR59) of porcine CD163 as porcine Fc (pFc) fusion proteins. Efficient expression of the soluble viral receptors in the rAd-transduced cells was confirmed by RT-PCR and Western blotting. To detect their antiviral activities, the soluble viral receptors were purified from the media of rAd-transduced cells and identified by Western blotting. The viral binding assay showed that the soluble receptors Sn4D-Fc and SRCR59-Fc, but not SRCR5-Fc and the control pFc, were able to bind to PRRSV particles. The viral infection blocking assays showed that co-treatment of PRRSV with different concentrations of Sn4D-Fc and SRCR59-Fc proteins resulted in a much higher (72.1%-77.6%) reduction in PRRSV-positive cell number than the single protein treatment (45.1%-60.0% or 44.0%-56.2%). To investigate the feasibility of delivering the soluble viral receptors to PAM, two pig cell lines were transduced with rAd-Sn4D-Fc and/or rAd-SRCR59-Fc using a transwell culture system. PAM cells were infected with PRRSV and then co-cultured with the rAd-transduced cells. Viral titration assay showed that co-cultivation of the infected PAM with rAd-Sn4D-Fc- and rAd-SRCR59-Fc-transduced cells resulted in much higher (by ?3.5 log) reduction in the viral titers (TCID50) than that of co-cultivation with the single vector-transduced cells (by ?1.0 log). Further studies showed that the rAd co-delivered soluble receptors Sn4D-Fc and SRCR59-Fc had dose-dependent and temporal antiviral effect against three different PRRSV strains. Since the data presented indicate an additive anti-PRRSV activity between the soluble receptors Sn4D-Fc and SRCR59-Fc, we conclude that the two rAd vectors generated will be useful for development a novel reagent for PRRS control. PMID:24246307

  4. No association found between the detection of either xenotropic murine leukemia virus-related virus or polytropic murine leukemia virus and chronic fatigue syndrome in a blinded, multi-site, prospective study by the establishment and use of the SolveCFS BioBank

    PubMed Central

    2014-01-01

    Background In 2009, a retrospective study reported the detection of xenotropic murine leukemia virus-related virus (XMRV) in clinical isolates derived from individuals with chronic fatigue syndrome or myalgic encephalomyelitis (CFS). While many efforts to confirm this observation failed, one report detected polytropic murine leukemia virus (pMLV), instead of XMRV. In both studies, Polymerase Chain Reaction (PCR)-based methods were employed which could provide the basis for the development of a practical diagnostic tool. To confirm these studies, we hypothesized that the ability to detect these viruses will not only depend upon the technical details of the methods employed but also on the criteria used to diagnose CFS and the availability of well characterized clinical isolates. Methods A repository of clinical isolates from geographically distinct sites was generated by the collection of fresh blood samples from well characterized CFS and healthy subjects. Molecular techniques were used to generate assay positive controls and to determine the lower limit of detection (LLOD) for murine retroviral and Intracisternal A particle (Cell 12(4):963-72, 1977) detection methods. Results We report the establishment of a repository of well-defined, clinical isolates from five, geographically distinct regions of the US, the comparative determination of the LLODs and validation efforts for the previously reported detection methods and the results of an effort to confirm the association of these retroviral signatures in isolates from individuals with CFS in a blinded, multi-site, prospective study. We detected various, murine retroviral DNA signatures but were unable to resolve a difference in the incidence of their detection between isolates from CFS (5/72; 6.7%) and healthy (2/37; 5.4%) subjects (Fisher’s Exact Test, p-value?=?1). The observed sequences appeared to reflect the detection of endogenous murine retroviral DNA, which was not identical to either XMRV or pMLV. Conclusions We were unable to confirm a previously reported association between the detection of XMRV or pMLV sequences and CFS in a prospective, multi-site study. Murine retroviral sequences were detected at a low frequency that did not differ between CFS and control subjects. The nature of these sequences appeared to reflect the detection of pre-existing, endogenous, murine retroviral DNA in the PCR reagents employed. PMID:25092471

  5. Metabolic Syndrome

    MedlinePLUS

    ... Your Best Self Smart Snacking Losing Weight Safely Metabolic Syndrome KidsHealth > Teens > Diabetes Center > Treatment & Prevention > Metabolic Syndrome ... applies to a condition known as metabolic syndrome. Metabolic Syndrome Is an Early Warning Sign Metabolic syndrome isn' ...

  6. 50 CFR 648.145 - Possession limit.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...one person, the violation shall be deemed to have been committed by the owner and operator. (d) Owners or operators of otter trawl vessels issued a moratorium permit under § 648.4 (a)(7) and fishing with, or possessing on board, nets or...

  7. 50 CFR 648.105 - Possession restrictions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...one person, the violation shall be deemed to have been committed by the owner and operator. (d) Owners and operators of otter trawl vessels issued a permit under § 648.4(a)(3) that fish with or possess nets or pieces of net on board that...

  8. 27 CFR 479.121 - Insular possessions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 3 2011-04-01 2010-04-01 true Insular possessions. 479.121 Section 479.121 Alcohol, Tobacco Products, and Firearms BUREAU OF ALCOHOL, TOBACCO, FIREARMS, AND EXPLOSIVES, DEPARTMENT OF JUSTICE FIREARMS AND AMMUNITION MACHINE GUNS, DESTRUCTIVE DEVICES, AND...

  9. 27 CFR 479.121 - Insular possessions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 3 2010-04-01 2010-04-01 false Insular possessions. 479.121 Section 479.121 Alcohol, Tobacco Products, and Firearms BUREAU OF ALCOHOL, TOBACCO, FIREARMS, AND EXPLOSIVES, DEPARTMENT OF JUSTICE FIREARMS AND AMMUNITION MACHINE GUNS, DESTRUCTIVE DEVICES, AND...

  10. 27 CFR 479.121 - Insular possessions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 3 2013-04-01 2013-04-01 false Insular possessions. 479.121 Section 479.121 Alcohol, Tobacco Products, and Firearms BUREAU OF ALCOHOL, TOBACCO, FIREARMS, AND EXPLOSIVES, DEPARTMENT OF JUSTICE FIREARMS AND AMMUNITION MACHINE GUNS, DESTRUCTIVE DEVICES, AND...

  11. 27 CFR 479.121 - Insular possessions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 3 2014-04-01 2014-04-01 false Insular possessions. 479.121 Section 479.121 Alcohol, Tobacco Products, and Firearms BUREAU OF ALCOHOL, TOBACCO, FIREARMS, AND EXPLOSIVES, DEPARTMENT OF JUSTICE FIREARMS AND AMMUNITION MACHINE GUNS, DESTRUCTIVE DEVICES, AND...

  12. 27 CFR 479.121 - Insular possessions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 3 2012-04-01 2010-04-01 true Insular possessions. 479.121 Section 479.121 Alcohol, Tobacco Products, and Firearms BUREAU OF ALCOHOL, TOBACCO, FIREARMS, AND EXPLOSIVES, DEPARTMENT OF JUSTICE FIREARMS AND AMMUNITION MACHINE GUNS, DESTRUCTIVE DEVICES, AND...

  13. Genome-Wide Gene Expression Profiles in Lung Tissues of Pig Breeds Differing in Resistance to Porcine Reproductive and Respiratory Syndrome Virus

    PubMed Central

    Zhang, Chenhua; Zhang, Yujie; Wang, Nan; Li, Yanping; Yang, Lijuan; Jiang, Chenglan; Zhang, Chaoyang; Wen, Changhong; Jiang, Yunliang

    2014-01-01

    Porcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus (PRRSV) is an infectious disease characterized by severe reproductive deficiency in pregnant sows, typical respiratory symptoms in piglets, and high mortality rate of piglets. In this study, we employed an Affymetrix microarray chip to compare the gene expression profiles of lung tissue samples from Dapulian (DPL) pigs (a Chinese indigenous pig breed) and Duroc×Landrace×Yorkshire (DLY) pigs after infection with PRRSV. During infection with PRRSV, the DLY pigs exhibited a range of clinical features that typify the disease, whereas the DPL pigs showed only mild signs of the disease. Overall, the DPL group had a lower percentage of CD4+ cells and lower CD4+/CD8+ratios than the DLY group (p<0.05). For both IL-10 and TNF-?, the DLY pigs had significantly higher levels than the DPL pigs (p<0.01). The DLY pigs have lower serum IFN-? levels than the DPL pigs (p<0.01). The serum IgG levels increased slightly from 0 dpi to 7 dpi, and peaked at 14 dpi (p<0.0001). Microarray data analysis revealed 16 differentially expressed (DE) genes in the lung tissue samples from the DLY and DPL pigs (q?5%), of which LOC100516029 and LOC100523005 were up-regulated in the PRRSV-infected DPL pigs, while the other 14 genes were down-regulated in the PRRSV-infected DPL pigs compared with the PRRSV-infected DLY pigs. The mRNA expression levels of 10 out of the 16 DE genes were validated by real-time quantitative RT-PCR and their fold change was consistent with the result of microarray data analysis. We further analyzed the mRNA expression level of 8 differentially expressed genes between the DPL and DLY pigs for both uninfected and infected groups, and found that TF and USP18 genes were important in underlying porcine resistance or susceptibility to PRRSV. PMID:24465897

  14. Influence of Agathi grandiflora active principles inhibit viral multiplication and stimulate immune system in Indian white shrimp Fenneropenaeus indicus against white spot syndrome virus infection.

    PubMed

    Bindhu, Francis; Velmurugan, Subramanian; Donio, Mariathason Birdilla Selva; Michaelbabu, Mariavincent; Citarasu, Thavasimuthu

    2014-12-01

    Five herbs including Adathoda vasica, Agathi grandiflora, Leucas aspera, Psoralea corylifolia, and Quercus infectoria were selected to screen the antiviral and immunostimulant activity against white spot syndrome virus (WSSV) and Vibrio harveyi respectively using different organic polar and non-polar solvents. Based on the initial screening results, ethyl acetate and methanolic extracts of A. grandiflora had strong antiviral and immunostimulant activities. Those extracts incubated with WSSV injected Fenneropenaeus indicus got only 20% mortality and no PCR positive signals were seen in two step PCR amplification. The methanolic extracts of A. grandiflora were further purified through silica column chromatography and the fractions screened again for antiviral and immunostimulant activity. The secondary screening results revealed that, the fractions of F5 to F7 had effectively controlled the WSSV multiplication and V. harveyi growth. The pooled fractions (F5 to F7) was structurally characterized by gas chromatograph-mass spectrometry (GC-MS) analysis and few compounds were identified including 3,7.11,15-Tetramethyl-2-Hexane-1-ol, pytol and 1,2-Benzenedicarboxylic acid, diisooctyl ester. The pooled fractions were mixed with the basal feed ingredients at the concentration of 100 (D-1), 200 (D-2), 300 (D-3) and 400 (D-4) mg kg(-1) and the diets fed to the F. indicus (9.0 ± 0.5 g) for 30 days. After the completion of feeding trail, they were challenged with virulent WSSV and studied the cumulative mortality, molecular diagnosis by quantitative real time PCR (qRT-PCR), biochemical, haematological and immunological parameters. The control diet fed F. indicus succumbed to death 100% within 3 days whereas the D-3 and D-4 helped to reduced the cumulative mortality of 60-80% respectively. The qRT-PCR revealed that, the WSSV copy number was gradually decreased when increasing concentration of A. grandiflora extract active fraction in the diets. The diets D-3 and D-4 helped to reduce the protein and carbohydrate levels significantly (P < 0.01) from the control diet fed groups. Moreover these diets help to decrease the coagulation time of maximum 61% from control groups and improve the total haemocyte count of maximum 51.82 × 10(5) cells ml(-1) in D4 diet fed F. indicus. Finally immunological parameters including prophenol oxidase (proPO) activity, intracellular superoxide anion production and intra-agar lysozyme activity was significantly (P ? 0.001) improved in the D-3 and D-4 fed F. indicus after WSSV challenge. PMID:25301717

  15. Acquired immunodeficiency syndrome/human immunodeficiency virus knowledge, attitudes, and practices, and use of healthcare services among rural migrants: a cross-sectional study in China

    PubMed Central

    2014-01-01

    Background Today’s rapid growth of migrant populations has been a major contributor to the human immunodeficiency virus (HIV) epidemic. However, relatively few studies have focused on HIV/acquired immunodeficiency syndrome (AIDS)-related knowledge, attitudes, and practice among rural-to-urban migrants in China. This cross-sectional study was to assess HIV/AIDS-related knowledge and perceptions, including knowledge about reducing high-risk sex. Methods Two-phase stratified cluster sampling was applied and 2,753 rural migrants participated in this study. An anonymous self-administered questionnaire was conducted in Guangdong and Sichuan provinces in 2007. Descriptive analysis was used to present the essential characteristics of the respondents. Chi-square test and multiple logistic regression models were performed to examine the associations between identified demographic factors and high-risk sex, sexually transmitted disease (STD) symptoms, and access to HIV screening services among the seven types of workers. Results 58.6% of participants were knowledgeable about HIV/AIDS transmission, but approximately 90% had a negative attitude towards the AIDS patients, and that 6.2% had engaged in high-risk sex in the past 12 months. Logistic regression analysis revealed sex, marital status, income, migration and work experience to be associated with high-risk sex. Among the 13.9% of workers who reported having STD symptoms, risk factors that were identified included female gender, high monthly income, being married, daily laborer or entertainment worker, frequent migration, and length of work experience. Only 3% of migrant workers received voluntary free HIV screening, which was positively associated with monthly income and workplace. Conclusions HIV/AIDS knowledge, attitudes, and practices among rural migrants in China remain a thorny health issue, and use of healthcare services needs to be improved. Low levels of education and knowledge regarding HIV/AIDS among housekeepers and migrant day laborers result in this population likely being engaged in high-risk sex. Government programs should pay more attention to public education, health promotion and intervention for the control of the HIV/AIDS epidemic in China. PMID:24520921

  16. Genome-wide gene expression profiles in lung tissues of pig breeds differing in resistance to porcine reproductive and respiratory syndrome virus.

    PubMed

    Xing, Jinyi; Xing, Feng; Zhang, Chenhua; Zhang, Yujie; Wang, Nan; Li, Yanping; Yang, Lijuan; Jiang, Chenglan; Zhang, Chaoyang; Wen, Changhong; Jiang, Yunliang

    2014-01-01

    Porcine reproductive and respiratory syndrome (PRRS) caused by PRRS virus (PRRSV) is an infectious disease characterized by severe reproductive deficiency in pregnant sows, typical respiratory symptoms in piglets, and high mortality rate of piglets. In this study, we employed an Affymetrix microarray chip to compare the gene expression profiles of lung tissue samples from Dapulian (DPL) pigs (a Chinese indigenous pig breed) and Duroc×Landrace×Yorkshire (DLY) pigs after infection with PRRSV. During infection with PRRSV, the DLY pigs exhibited a range of clinical features that typify the disease, whereas the DPL pigs showed only mild signs of the disease. Overall, the DPL group had a lower percentage of CD4(+) cells and lower CD4(+)/CD8(+)ratios than the DLY group (p<0.05). For both IL-10 and TNF-?, the DLY pigs had significantly higher levels than the DPL pigs (p<0.01). The DLY pigs have lower serum IFN-? levels than the DPL pigs (p<0.01). The serum IgG levels increased slightly from 0 dpi to 7 dpi, and peaked at 14 dpi (p<0.0001). Microarray data analysis revealed 16 differentially expressed (DE) genes in the lung tissue samples from the DLY and DPL pigs (q?5%), of which LOC100516029 and LOC100523005 were up-regulated in the PRRSV-infected DPL pigs, while the other 14 genes were down-regulated in the PRRSV-infected DPL pigs compared with the PRRSV-infected DLY pigs. The mRNA expression levels of 10 out of the 16 DE genes were validated by real-time quantitative RT-PCR and their fold change was consistent with the result of microarray data analysis. We further analyzed the mRNA expression level of 8 differentially expressed genes between the DPL and DLY pigs for both uninfected and infected groups, and found that TF and USP18 genes were important in underlying porcine resistance or susceptibility to PRRSV. PMID:24465897

  17. Biogeographic and Ecological Regulation of Disease: Prevalence of Sin Nombre Virus in Island Mice Is Related to Island Area, Precipitation, and Predator Richness.

    E-print Network

    Allan, Brian

    virus (SNV), the agent of a severe disease in humans (hantavirus pulmonary syndrome), in island deer Channel Islands, disease ecology, habitat area, hantavirus, predators, Sin Nombre virus. Introduction

  18. Dynamics of Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2) DNA loads in serum of healthy and postweaning multisystemic wasting syndrome (PMWS) affected pigs.

    PubMed

    Nieto, D; Aramouni, M; Grau-Roma, L; Segalés, J; Kekarainen, T

    2011-09-28

    Torque teno viruses (TTVs) are vertebrate infecting, small viruses with circular single stranded DNA, classified in the Anelloviridae family. In pigs, two different TTV species have been described so far, Torque teno sus virus 1 (TTSuV1) and 2 (TTSuV2). TTSuVs have lately been linked to postweaning multisystemic wasting syndrome (PMWS). In the present study, TTSuV1 and TTSuV2 prevalence and DNA loads in longitudinally collected serum samples of healthy and PMWS affected pigs from Spanish conventional, multi-site farms were analyzed. Serum samples were taken at 1, 3, 7, 11 and around 15 weeks of age (age of PMWS outbreak) and viral DNA loads determined by quantitative PCR. For both TTSuV species, percentage of viremic pigs increased progressively over time, with the highest prevalence in animals of about 15 weeks of age. TTSuV1 and TTSuV2 viral DNA loads in healthy and TTSuV1 loads in PMWS affected animals increased until 11 weeks of age declining afterwards. On the contrary, TTSuV2 DNA loads in PMWS affected pigs increased throughout the sampling period. It seems that TTSuV species differ in the in vivo infection dynamics in PMWS affected animals. PMID:21680113

  19. Possession Versus Position: Strategic Evaluation in AFL

    PubMed Central

    O’Shaughnessy, Darren M.

    2006-01-01

    In sports like Australian Rules football and soccer, teams must battle to achieve possession of the ball in sufficient space to make optimal use of it. Ultimately the teams need to score, and to do that the ball must be brought into the area in front of goal - the place where the defence usually concentrates on shutting down space and opportunity time. Coaches would like to quantify the trade-offs between contested play in good positions and uncontested play in less promising positions, in order to inform their decision-making about where to put their players, and when to gamble on sending the ball to a contest rather than simply maintain possession. To evaluate football strategies, Champion Data has collected the on-ground locations of all 350,000 possessions and stoppages in the past two seasons of AFL (2004, 2005). By following each chain of play through to the next score, we can now reliably estimate the scoreboard “equity ”of possessing the ball at any location, and measure the effect of having sufficient time to dispose of it effectively. As expected, winning the ball under physical pressure (through a “hard ball get”) is far more difficult to convert into a score than winning it via a mark. We also analyse some equity gradients to show how getting the ball 20 metres closer to goal is much more important in certain areas of the ground than in others. We conclude by looking at the choices faced by players in possession wanting to maximise their likelihood of success. Key Points Equity analysis provides a way of estimating the net value of actions on the sporting field. Combined with spatial data analysis, the relative merits of gaining position or maintaining possession can be judged. The advantage of having time and space to use the ball is measured in terms of scoreboard value, and is found to vary with field position. Each sport has identifiable areas of the field with high equity gradients, meaning that it is most important to gain territory there. PMID:24357947

  20. Does Rhodiola rosea possess ergogenic properties?

    PubMed

    Walker, Thomas B; Robergs, Robert A

    2006-06-01

    Rhodiola rosea is an herb purported to possess adaptogenic and ergogenic properties and has recently been the subject of increased interest The purpose of this article was to review and summarize recent investigations of the potential performance-enhancing properties of Rhodiola rosea. Such studies have generated equivocal results. Several investigations conducted in Eastern Europe have indicated that Rhodiola rosea ingestion may produce such positive effects as improved cognitive function and reduced mental fatigue. Other research from this region has illustrated enhanced endurance exercise performance in both humans and rats. Studies conducted in Western Europe and in North America have indicated that Rhodiola rosea may possess substantial antioxidant properties but have produced mixed results when attempting to demonstrate an ergogenic effect during exercise in humans. PMID:16948486

  1. Tracking the actions and possessions of agents

    PubMed Central

    Gelman, Susan A.; Noles, Nicholaus S.; Stilwell, Sarah

    2014-01-01

    We propose that there is a powerful human disposition to track the actions and possessions of agents. In two experiments, 3-year-olds and adults viewed sets of objects, learned a new fact about one of the objects in each set (either that it belonged to the participant, or that it possessed a particular label), and were queried about either the taught fact or an unrelated dimension (preference) immediately after a spatiotemporal transformation, and after a delay. Adults uniformly tracked object identity under all conditions, whereas children tracked identity more when taught ownership versus labeling information, and only regarding the taught fact (not the unrelated dimension). These findings suggest that the special attention that children and adults pay to agents readily extends to include inanimate objects. That young children track an object’s history, despite their reliance on surface features on many cognitive tasks, suggests that unobservable historical features are foundational in human cognition. PMID:25111732

  2. 25 CFR 11.452 - Possession of a controlled substance.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... false Possession of a controlled substance. 11.452 Section 11...11.452 Possession of a controlled substance. (a) It is unlawful...intentionally possess any controlled substance listed in 21 CFR Part...

  3. 25 CFR 11.452 - Possession of a controlled substance.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... false Possession of a controlled substance. 11.452 Section 11...11.452 Possession of a controlled substance. (a) It is unlawful...intentionally possess any controlled substance listed in 21 CFR Part...

  4. 25 CFR 11.452 - Possession of a controlled substance.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... false Possession of a controlled substance. 11.452 Section 11...11.452 Possession of a controlled substance. (a) It is unlawful...intentionally possess any controlled substance listed in 21 CFR Part...

  5. 25 CFR 11.452 - Possession of a controlled substance.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... false Possession of a controlled substance. 11.452 Section 11...11.452 Possession of a controlled substance. (a) It is unlawful...intentionally possess any controlled substance listed in 21 CFR Part...

  6. 25 CFR 11.452 - Possession of a controlled substance.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... true Possession of a controlled substance. 11.452 Section 11...11.452 Possession of a controlled substance. (a) It is unlawful...intentionally possess any controlled substance listed in 21 CFR Part...

  7. Clinical Outcome and Genetic Differences within a Monophyletic Dengue Virus Type 2 Population

    PubMed Central

    Shi, Yuan; Thein, Tun Lin; Lee, Linda Kay; Lee, Kim Sung; Lye, David Chien; Ng, Lee Ching; Leo, Yee Sin

    2015-01-01

    The exact mechanisms of interplay between host and viral factors leading to severe dengue are yet to be fully understood. Even though previous studies have implicated specific genetic differences of Dengue virus (DENV) in clinical severity and virus attenuation, similar studies with large-scale, whole genome screening of monophyletic virus populations are limited. Therefore, in the present study, we compared 89 whole genomes of DENV-2 cosmopolitan clade III isolates obtained from patients diagnosed with dengue fever (DF, n = 58), dengue hemorrhagic fever (DHF, n = 30) and dengue shock syndrome (DSS, n = 1) in Singapore between July 2010 and January 2013, in order to determine the correlation of observed viral genetic differences with clinical outcomes. Our findings showed no significant difference between the number of primary and secondary infections that progressed to DHF and DSS (p>0.05) in our study cohort. Despite being highly homogenous, study isolates possessed 39 amino acid substitutions of which 10 substitutions were fixed in three main groups of virus isolates. None of those substitutions were specifically associated with DHF and DSS. Notably, two evolutionarily unique virus groups possessing C-P43T+NS1-S103T+NS2A-V83I+NS3-R337K+ NS3-I600T+ NS5-P136S and NS2A-T119N mutations were exclusively found in patients with DF, the benign form of DENV infections. Those mutants were significantly associated with mild disease outcome. These observations indicated that disease progression into DHF and DSS within our patient population was more likely to be due to host than virus factors. We hypothesize that selection for potentially less virulent groups of DENV-2 in our study cohort may be an evolutionary adaptation of viral strains to extend their survival in the human-mosquito transmission cycle. PMID:25811657

  8. Epstein-Barr virus-related post-transplant lymphoproliferative disorder occurring after bone marrow transplantation for aplastic anemia in Down's syndrome.

    PubMed

    Furuya, Aya; Ishida, Mitsuaki; Hodohara, Keiko; Yoshii, Miyuki; Okuno, Hiroko; Horinouchi, Akiko; Nakanishi, Ryota; Harada, Ayumi; Iwai, Muneo; Yoshida, Keiko; Kagotani, Akiko; Yoshida, Takashi; Okabe, Hidetoshi

    2014-01-01

    It is well established that Down's syndrome exhibits a predisposition to development of leukemia, however, association between aplastic anemia and Down's syndrome is exceptional. Herein, we describe a case of aplastic anemia occurring in Down's syndrome following post-transplant lymphoproliferative disorder (PTLD) after bone marrow transplantation (BMT). A 27-year-old Japanese male with Down's syndrome presented with a headache. Laboratory tests revealed severe pancytopenia, and bone marrow biopsy demonstrated hypocellular bone marrow with decrease of trilineage cells, which led to a diagnosis of aplastic anemia. One year after diagnosis, he was incidentally found to have an anterior mediastinal tumor, which was histopathologically diagnosed as seminoma. Subsequently, he received BMT from a female donor, and engraftment was observed. Three months after transplantation, he experienced cough and high fever. Biopsy specimen from the lung revealed diffuse proliferation of large-sized lymphoid cells expressing CD20 and EBER. These lymphoid cells had XY chromosomes. Thus, a diagnosis of EBV-associated PTLD was made. This is the seventh documented case of aplastic anemia occurring in Down's syndrome. Association between aplastic anemia and Down's syndrome has not been established, therefore, additional clinicopathological studies are needed. Moreover, this is the first case to undergo BMT for aplastic anemia in Down's syndrome. Although engraftment was observed, he developed EBV-positive PTLD. The neoplastic cells of the present case were considered to be of recipient origin, although the majority of PTLD cases with BMT are of donor origin. PMID:24427369

  9. Bat Flight and Zoonotic Viruses

    PubMed Central

    Cryan, Paul M.; Cunningham, Andrew A.; Fooks, Anthony R.; Hayman, David T.S.; Luis, Angela D.; Peel, Alison J.; Plowright, Raina K.; Wood, James L.N.

    2014-01-01

    Bats are sources of high viral diversity and high-profile zoonotic viruses worldwide. Although apparently not pathogenic in their reservoir hosts, some viruses from bats severely affect other mammals, including humans. Examples include severe acute respiratory syndrome coronaviruses, Ebola and Marburg viruses, and Nipah and Hendra viruses. Factors underlying high viral diversity in bats are the subject of speculation. We hypothesize that flight, a factor common to all bats but to no other mammals, provides an intensive selective force for coexistence with viral parasites through a daily cycle that elevates metabolism and body temperature analogous to the febrile response in other mammals. On an evolutionary scale, this host–virus interaction might have resulted in the large diversity of zoonotic viruses in bats, possibly through bat viruses adapting to be more tolerant of the fever response and less virulent to their natural hosts. PMID:24750692

  10. Bat flight and zoonotic viruses

    USGS Publications Warehouse

    O'Shea, Thomas; Cryan, Paul M.; Cunningham, Andrew A.; Fooks, Anthony R.; Hayman, David T.S.; Luis, Angela D.; Peel, Alison J.; Plowright, Raina K.; Wood, James L.N.

    2014-01-01

    Bats are sources of high viral diversity and high-profile zoonotic viruses worldwide. Although apparently not pathogenic in their reservoir hosts, some viruses from bats severely affect other mammals, including humans. Examples include severe acute respiratory syndrome coronaviruses, Ebola and Marburg viruses, and Nipah and Hendra viruses. Factors underlying high viral diversity in bats are the subject of speculation. We hypothesize that flight, a factor common to all bats but to no other mammals, provides an intensive selective force for coexistence with viral parasites through a daily cycle that elevates metabolism and body temperature analogous to the febrile response in other mammals. On an evolutionary scale, this host–virus interaction might have resulted in the large diversity of zoonotic viruses in bats, possibly through bat viruses adapting to be more tolerant of the fever response and less virulent to their natural hosts.

  11. Metabolic Syndrome

    MedlinePLUS

    ... Th M e etabolic Syndrome What is the metabolic syndrome? The term metabolic syndrome describes a cluster of risk factors that increase ... high blood sugar). The exact cause of the metabolic syndrome is not known but genetic factors, too much ...

  12. Cushing's Syndrome

    MedlinePLUS

    MENU Return to Web version Cushing's Syndrome Overview What is Cushing's syndrome? Cushing's syndrome occurs when your body is exposed to high levels ... they can cause problems with your eyesight. Diagnosis & Tests How is Cushing's syndrome diagnosed? Your doctor may ...

  13. Expression of Ley antigen in human immunodeficiency virus-infected human T cell lines and in peripheral lymphocytes of patients with acquired immune deficiency syndrome (AIDS) and AIDS-related complex (ARC).

    PubMed

    Adachi, M; Hayami, M; Kashiwagi, N; Mizuta, T; Ohta, Y; Gill, M J; Matheson, D S; Tamaoki, T; Shiozawa, C; Hakomori, S

    1988-02-01

    Ley determinant (Fuc alpha 1----2Gal beta 1----4[Fuc alpha 1----3]GlcNAc beta 1----R) defined by mAb BM-1 is highly expressed in human immunodeficiency virus (HIV)-infected T cell lines and in CD3+ peripheral mature T cells of patients with acquired immune deficiency syndrome (AIDS) or with AIDS-related complex (ARC). Ley expression increased greatly in the CD3+ population in the advanced stage of AIDS when the CD4+ population decreased greatly. Six other carbohydrate antigens tested by their respective mAbs were not detected in these same cells. None of the carbohydrate antigens tested by the seven mAbs used in this study were found in noninfected T cell lines and in normal peripheral blood lymphocytes. PMID:3258005

  14. 50 CFR 20.38 - Possession of live birds.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 6 2010-10-01 2010-10-01 false Possession of live birds. 20.38 Section 20... WILDLIFE AND PLANTS (CONTINUED) MIGRATORY BIRD HUNTING Possession § 20.38 Possession of live birds. Every migratory game bird wounded by hunting and reduced to possession by the hunter shall be immediately...

  15. 50 CFR 20.38 - Possession of live birds.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 50 Wildlife and Fisheries 8 2011-10-01 2011-10-01 false Possession of live birds. 20.38 Section 20... WILDLIFE AND PLANTS (CONTINUED) MIGRATORY BIRD HUNTING Possession § 20.38 Possession of live birds. Every migratory game bird wounded by hunting and reduced to possession by the hunter shall be immediately...

  16. 50 CFR 20.38 - Possession of live birds.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 9 2013-10-01 2013-10-01 false Possession of live birds. 20.38 Section 20... WILDLIFE AND PLANTS (CONTINUED) MIGRATORY BIRD HUNTING Possession § 20.38 Possession of live birds. Every migratory game bird wounded by hunting and reduced to possession by the hunter shall be immediately...

  17. 50 CFR 20.38 - Possession of live birds.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 9 2012-10-01 2012-10-01 false Possession of live birds. 20.38 Section 20... WILDLIFE AND PLANTS (CONTINUED) MIGRATORY BIRD HUNTING Possession § 20.38 Possession of live birds. Every migratory game bird wounded by hunting and reduced to possession by the hunter shall be immediately...

  18. 50 CFR 20.38 - Possession of live birds.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 9 2014-10-01 2014-10-01 false Possession of live birds. 20.38 Section 20... WILDLIFE AND PLANTS (CONTINUED) MIGRATORY BIRD HUNTING Possession § 20.38 Possession of live birds. Every migratory game bird wounded by hunting and reduced to possession by the hunter shall be immediately...

  19. Epstein-Barr virus-related post-transplant lymphoproliferative disorder occurring after bone marrow transplantation for aplastic anemia in Down’s syndrome

    PubMed Central

    Furuya, Aya; Ishida, Mitsuaki; Hodohara, Keiko; Yoshii, Miyuki; Okuno, Hiroko; Horinouchi, Akiko; Nakanishi, Ryota; Harada, Ayumi; Iwai, Muneo; Yoshida, Keiko; Kagotani, Akiko; Yoshida, Takashi; Okabe, Hidetoshi

    2014-01-01

    It is well established that Down’s syndrome exhibits a predisposition to development of leukemia, however, association between aplastic anemia and Down’s syndrome is exceptional. Herein, we describe a case of aplastic anemia occurring in Down’s syndrome following post-transplant lymphoproliferative disorder (PTLD) after bone marrow transplantation (BMT). A 27-year-old Japanese male with Down’s syndrome presented with a headache. Laboratory tests revealed severe pancytopenia, and bone marrow biopsy demonstrated hypocellular bone marrow with decrease of trilineage cells, which led to a diagnosis of aplastic anemia. One year after diagnosis, he was incidentally found to have an anterior mediastinal tumor, which was histopathologically diagnosed as seminoma. Subsequently, he received BMT from a female donor, and engraftment was observed. Three months after transplantation, he experienced cough and high fever. Biopsy specimen from the lung revealed diffuse proliferation of large-sized lymphoid cells expressing CD20 and EBER. These lymphoid cells had XY chromosomes. Thus, a diagnosis of EBV-associated PTLD was made. This is the seventh documented case of aplastic anemia occurring in Down’s syndrome. Association between aplastic anemia and Down’s syndrome has not been established, therefore, additional clinicopathological studies are needed. Moreover, this is the first case to undergo BMT for aplastic anemia in Down’s syndrome. Although engraftment was observed, he developed EBV-positive PTLD. The neoplastic cells of the present case were considered to be of recipient origin, although the majority of PTLD cases with BMT are of donor origin. PMID:24427369

  20. A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China.

    PubMed

    Chai, Zheng; Ma, Wenjun; Fu, Fang; Lang, Yuekun; Wang, Wei; Tong, Guangzhi; Liu, Qinfang; Cai, Xuehui; Li, Xi

    2013-02-01

    SYBR Green coupled to melting curve analysis has been suggested to detect RNA viruses showing high genomic variability. Here, a SYBR Green-based real-time RT-PCR assay was developed for simultaneous detection and differentiation of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) and classical type 2 PRRSV (C-PRRSV). The different strains were identified by their distinctive melting temperatures: 82.98 ± 0.25 °C and 85.95 ± 0.24 °C for HP-PRRSVs or 82.74 ± 0.26 °C for C-PRRSVs. Specificity was tested using nine other viral and bacterial pathogens of swine. The detection limit was 1 TCID(50) for HP- or C-PRRSV. Furthermore, the detection results for samples from an animal trial with HP- or C-PRRSV infections showed that the SYBR Green-based real-time RT-PCR was more sensitive than the conventional RT-PCR. Additionally, an analysis of 319 field samples from North China, Central China and Northeast China showed that HP- and C-PRRSVs co-circulated in pig herds. Thus, the SYBR Green-based real-time RT-PCR, which can be performed within one hour, is a rapid, sensitive and low-cost diagnostic tool for rapid differential detection and routine surveillance of HP- and classical type 2 PRRSVs in China. PMID:23070137

  1. Phase transitions in systems possessing shock solutions

    NASA Astrophysics Data System (ADS)

    Arabsalmani, Maryam; Aghamohammadi, Amir

    2006-07-01

    Recently it has been shown that there are three families of stochastic one-dimensional nonequilibrium lattice models for which the single-shock measures form an invariant subspace of the states of these models. Here, both the stationary states and dynamics of single-shocks on a one-dimensional lattice are studied. This is done for both an infinite lattice and a finite lattice with boundaries. It is seen that these models possess both static and dynamical phase transitions. The static phase transition is the well-known low-high density phase transition for the asymmetric simple exclusion process. The branching-coalescing random walk and asymmetric Kawasaki-Glauber process models also show the same phase transition. Double-shocks on a one-dimensional lattice are also investigated. It is shown that at the stationary state the contribution of double-shocks with higher width becomes small, and the main contribution comes from thin double-shocks.

  2. Planctomycetes do possess a peptidoglycan cell wall

    PubMed Central

    Jeske, Olga; Schüler, Margarete; Schumann, Peter; Schneider, Alexander; Boedeker, Christian; Jogler, Mareike; Bollschweiler, Daniel; Rohde, Manfred; Mayer, Christoph; Engelhardt, Harald; Spring, Stefan; Jogler, Christian

    2015-01-01

    Most bacteria contain a peptidoglycan (PG) cell wall, which is critical for maintenance of shape and important for cell division. In contrast, Planctomycetes have been proposed to produce a proteinaceous cell wall devoid of PG. The apparent absence of PG has been used as an argument for the putative planctomycetal ancestry of all bacterial lineages. Here we show, employing multiple bioinformatic methods, that planctomycetal genomes encode proteins required for PG synthesis. Furthermore, we biochemically demonstrate the presence of the sugar and the peptide components of PG in Planctomycetes. In addition, light and electron microscopic experiments reveal planctomycetal PG sacculi that are susceptible to lysozyme treatment. Finally, cryo-electron tomography demonstrates that Planctomycetes possess a typical PG cell wall and that their cellular architecture is thus more similar to that of other Gram-negative bacteria. Our findings shed new light on the cellular architecture and cell division of the maverick Planctomycetes. PMID:25964217

  3. A fast and robust method for full genome sequencing of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Type 1 and Type 2.

    PubMed

    Kvisgaard, Lise K; Hjulsager, Charlotte K; Fahnøe, Ulrik; Breum, Solvej Ø; Ait-Ali, Tahar; Larsen, Lars E

    2013-11-01

    PRRSV is a positive-sense RNA virus with a high degree of genetic variability among isolates. For diagnostic sensitivity and vaccine design it is essential to monitor PRRSV genetic diversity. However, to date only a few full genome sequences of PRRSV isolates have been made publicly available. In the present study, fast and robust methods for long range RT-PCR amplification and subsequent next generation sequencing (NGS) were developed and validated on nine Type 1 and nine Type 2 PRRSV viruses. The methods generated robust and reliable sequences both on primary material and cell culture adapted viruses and the protocols performed well on all three NGS platforms tested (Roche 454 FLX, Illumina HiSeq2000, and Ion Torrent PGM™ Sequencer). These methods will greatly facilitate the generation of more full genome PRRSV sequences globally. PMID:23891870

  4. Azidothymidine triphosphate is an inhibitor of both human immunodeficiency virus type 1 reverse transcriptase and DNA polymerase gamma.

    PubMed Central

    König, H; Behr, E; Löwer, J; Kurth, R

    1989-01-01

    The reverse transcriptase from human immunodeficiency virus type 1 was purified from the virus to near homogeneity. The enzyme was shown to possess both RNA-dependent and DNA-dependent DNA-synthesizing activity. Activated DNA as a heteropolymeric substrate was used as efficiently as was the homopolymeric substrate poly(rA)-oligo(dT). The Michaelis-Menten constants were determined for each of the four nucleotides needed to elongate a natural template primer. Azidothymidine triphosphate, a well-known inhibitor of the enzyme, inhibited the enzyme competitively with respect to dTTP and noncompetitively with respect to the other nucleotides. Azidothymidine triphosphate acted as an efficient inhibitor of cellular DNA polymerase gamma, whereas other enzymes of eucaryotic DNA metabolism, namely, DNA polymerase alpha-primase and DNA polymerase beta, were not inhibited. This finding may explain why some acquired immunodeficiency syndrome patients suffer side effects during azidothymidine therapy. Images PMID:2482702

  5. The pathogenesis of foot-and-mouth disease II; viral pathways in swine, small ruminants, and wildlife, myotropism, chronic syndromes, and molecular virus-host interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Investigation of the pathogenesis of foot-and-mouth disease (FMD) has focused on study of the disease in cattle with less emphasis on pigs, small ruminants, and wildlife. “Atypical” FMD-associated syndromes such as myocarditis, reproductive losses, and chronic heat-intolerance have also received lit...

  6. Phase transitions in systems possessing shock solutions.

    PubMed

    Arabsalmani, Maryam; Aghamohammadi, Amir

    2006-07-01

    Recently it has been shown that there are three families of stochastic one-dimensional nonequilibrium lattice models for which the single-shock measures form an invariant subspace of the states of these models. Here, both the stationary states and dynamics of single-shocks on a one-dimensional lattice are studied. This is done for both an infinite lattice and a finite lattice with boundaries. It is seen that these models possess both static and dynamical phase transitions. The static phase transition is the well-known low-high density phase transition for the asymmetric simple exclusion process. The branching-coalescing random walk and asymmetric Kawasaki-Glauber process models also show the same phase transition. Double-shocks on a one-dimensional lattice are also investigated. It is shown that at the stationary state the contribution of double-shocks with higher width becomes small, and the main contribution comes from thin double-shocks. PMID:16907060

  7. RECQ1 possesses DNA branch migration activity.

    PubMed

    Bugreev, Dmitry V; Brosh, Robert M; Mazin, Alexander V

    2008-07-18

    RecQ helicases are essential for the maintenance of genome stability. Five members of the RecQ family have been found in humans, including RECQ1, RECQ5, BLM, WRN, and RECQ4; the last three are associated with human diseases. At this time, only BLM and WRN helicases have been extensively characterized, and the information on the other RecQ helicases has only started to emerge. Our current paper is focused on the biochemical properties of human RECQ1 helicase. Recent cellular studies have shown that RECQ1 may participate in DNA repair and homologous recombination, but the exact mechanisms of how RECQ1 performs its cellular functions remain largely unknown. Whereas RECQ1 possesses poor helicase activity, we found here that the enzyme efficiently promotes DNA branch migration. Further analysis revealed that RECQ1 catalyzes unidirectional three-stranded branch migration with a 3' --> 5' polarity. We show that this RECQ1 activity is instrumental in specific disruption of joint molecules (D-loops) formed by a 5' single-stranded DNA invading strand, which may represent dead end intermediates of homologous recombination in vivo. The newly found enzymatic properties of the RECQ1 helicase may have important implications for the function of RECQ1 in maintenance of genomic stability. PMID:18495662

  8. Propidium monoazide (PMA) and ethidium bromide monoazide (EMA) improve DNA array and high-throughput sequencing of porcine reproductive and respiratory syndrome virus identification.

    PubMed

    Bellehumeur, Christian; Boyle, Brian; Charette, Steve J; Harel, Josée; L'Homme, Yvan; Masson, Luke; Gagnon, Carl A

    2015-09-15

    Pan-viral DNA array (PVDA) and high-throughput sequencing (HTS) are useful tools to identify novel viruses of emerging diseases. However, both techniques have difficulties to identify viruses in clinical samples because of the host genomic nucleic acid content (hg/cont). Both propidium monoazide (PMA) and ethidium bromide monoazide (EMA) have the capacity to bind free DNA/RNA, but are cell membrane-impermeable. Thus, both are unable to bind protected nucleic acid such as viral genomes within intact virions. However, EMA/PMA modified genetic material cannot be amplified by enzymes. In order to assess the potential of EMA/PMA to lower the presence of amplifiable hg/cont in samples and improve virus detection, serum and lung tissue homogenates were spiked with porcine reproductive and respiratory virus (PRRSV) and were processed with EMA/PMA. In addition, PRRSV RT-qPCR positive clinical samples were also tested. EMA/PMA treatments significantly decreased amplifiable hg/cont and significantly increased the number of PVDA positive probes and their signal intensity compared to untreated spiked lung samples. EMA/PMA treatments also increased the sensitivity of HTS by increasing the number of specific PRRSV reads and the PRRSV percentage of coverage. Interestingly, EMA/PMA treatments significantly increased the sensitivity of PVDA and HTS in two out of three clinical tissue samples. Thus, EMA/PMA treatments offer a new approach to lower the amplifiable hg/cont in clinical samples and increase the success of PVDA and HTS to identify viruses. PMID:26129867

  9. Hyperferritinemic syndrome: Still's disease and catastrophic antiphospholipid syndrome triggered by fulminant Chikungunya infection: a case report of two patients.

    PubMed

    Betancur, Juan-Felipe; Navarro, Erika-Paola; Echeverry, Alex; Moncada, Pablo A; Cañas, Carlos A; Tobón, Gabriel J

    2015-11-01

    There are four medical conditions characterized by high levels of ferritin, the macrophage activation syndrome (MAS), adult onset Still' s disease (AOSD), catastrophic antiphospholipid syndrome (CAPS), and septic shock, that share similar clinical and laboratory features, suggesting a common pathogenic mechanism. This common syndrome entity is termed "the hyperferritinemic syndrome." Here, we describe two different cases of hyperferritinemic syndrome triggered by Chikungunya fever virus infection: a 21-year-old female with SLE and a 32-year-old male patient who developed AOSD after the coinfection of dengue and Chikungunya viruses. PMID:26233722

  10. 31 CFR 0.215 - Possession of weapons and explosives.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...2010-07-01 false Possession of weapons and explosives. 0.215 Section 0...of Conduct § 0.215 Possession of weapons and explosives. (a) Employees shall...explosives, or other dangerous or deadly weapons, either openly or concealed,...

  11. 50 CFR 622.277 - Bag and possession limits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...GULF OF MEXICO, AND SOUTH ATLANTIC Dolphin and Wahoo Fishery Off the Atlantic States...possession limits. (a) Atlantic dolphin and wahoo. Bag and possession limits are as follows: (1) Dolphin—10, not to exceed 60 per...

  12. 50 CFR 622.277 - Bag and possession limits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...GULF OF MEXICO, AND SOUTH ATLANTIC Dolphin and Wahoo Fishery Off the Atlantic States...possession limits. (a) Atlantic dolphin and wahoo. Bag and possession limits are as follows: (1) Dolphin—10, not to exceed 60 per...

  13. 50 CFR 648.235 - Possession and landing restrictions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 8 2010-10-01 2010-10-01 false Possession and landing restrictions. 648... Measures for the Spiny Dogfish Fishery § 648.235 Possession and landing restrictions. (a) Quota Period 1... calendar day. (c) Regulations governing the harvest, possession, landing, purchase, and sale of shark...

  14. 50 CFR 648.52 - Possession and landing limits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 50 Wildlife and Fisheries 8 2010-10-01 2010-10-01 false Possession and landing limits. 648.52... Measures for the Atlantic Sea Scallop Fishery § 648.52 Possession and landing limits. (a) A vessel issued... Program as described in § 648.60 are prohibited from fishing for or landing per trip, or possessing at...

  15. 31 CFR 0.215 - Possession of weapons and explosives.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 31 Money and Finance: Treasury 1 2014-07-01 2014-07-01 false Possession of weapons and explosives... OF THE TREASURY EMPLOYEE RULES OF CONDUCT Rules of Conduct § 0.215 Possession of weapons and explosives. (a) Employees shall not possess firearms, explosives, or other dangerous or deadly...

  16. 31 CFR 0.215 - Possession of weapons and explosives.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 31 Money and Finance: Treasury 1 2010-07-01 2010-07-01 false Possession of weapons and explosives... OF THE TREASURY EMPLOYEE RULES OF CONDUCT Rules of Conduct § 0.215 Possession of weapons and explosives. (a) Employees shall not possess firearms, explosives, or other dangerous or deadly...

  17. 31 CFR 0.215 - Possession of weapons and explosives.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 31 Money and Finance: Treasury 1 2012-07-01 2012-07-01 false Possession of weapons and explosives... OF THE TREASURY EMPLOYEE RULES OF CONDUCT Rules of Conduct § 0.215 Possession of weapons and explosives. (a) Employees shall not possess firearms, explosives, or other dangerous or deadly...

  18. 31 CFR 0.215 - Possession of weapons and explosives.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 31 Money and Finance: Treasury 1 2011-07-01 2011-07-01 false Possession of weapons and explosives... OF THE TREASURY EMPLOYEE RULES OF CONDUCT Rules of Conduct § 0.215 Possession of weapons and explosives. (a) Employees shall not possess firearms, explosives, or other dangerous or deadly...

  19. 31 CFR 0.215 - Possession of weapons and explosives.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 31 Money and Finance: Treasury 1 2013-07-01 2013-07-01 false Possession of weapons and explosives... explosives. (a) Employees shall not possess firearms, explosives, or other dangerous or deadly weapons... paragraph (a) of this section does not apply to employees who are required to possess weapons or...

  20. 50 CFR 622.187 - Bag and possession limits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...2013-10-01 2013-10-01 false Bag and possession limits. 622.187 Section...the South Atlantic Region § 622.187 Bag and possession limits. (a) Additional...provides the general applicability for bag and possession limits. However, §...

  1. 50 CFR 622.187 - Bag and possession limits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...2014-10-01 2014-10-01 false Bag and possession limits. 622.187 Section...the South Atlantic Region § 622.187 Bag and possession limits. (a) Additional...provides the general applicability for bag and possession limits. However, §...

  2. 50 CFR 622.38 - Bag and possession limits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...2014-10-01 2014-10-01 false Bag and possession limits. 622.38 Section...Resources of the Gulf of Mexico § 622.38 Bag and possession limits. (a) Additional...provides the general applicability for bag and possession limits. However, §...

  3. 50 CFR 622.38 - Bag and possession limits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...2013-10-01 2013-10-01 false Bag and possession limits. 622.38 Section...Resources of the Gulf of Mexico § 622.38 Bag and possession limits. (a) Additional...provides the general applicability for bag and possession limits. However, §...

  4. 50 CFR 648.94 - Monkfish possession and landing restrictions.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...determined by multiplying the tail weight by 1.91. For example a vessel possessing 100 lb (45 kg) of tail weight may possess an additional 191 lb...heads only without possessing the equivalent weight of tails allowed by using the...

  5. 50 CFR 648.94 - Monkfish possession and landing restrictions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...determined by multiplying the tail weight by 1.91. For example a vessel possessing 100 lb (45 kg) of tail weight may possess an additional 191 lb...heads only without possessing the equivalent weight of tails allowed by using the...

  6. 50 CFR 648.94 - Monkfish possession and landing restrictions.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...determined by multiplying the tail weight by 1.91. For example a vessel possessing 100 lb (45 kg) of tail weight may possess an additional 191 lb...heads only without possessing the equivalent weight of tails allowed by using the...

  7. 50 CFR 648.94 - Monkfish possession and landing restrictions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...determined by multiplying the tail weight by 1.91. For example a vessel possessing 100 lb (45 kg) of tail weight may possess an additional 191 lb...heads only without possessing the equivalent weight of tails allowed by using the...

  8. 50 CFR 648.145 - Black sea bass possession limit.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 50 Wildlife and Fisheries 12 2012-10-01 2012-10-01 false Black sea bass possession limit. 648.145... Measures for the Black Sea Bass Fishery § 648.145 Black sea bass possession limit. (a) From January 1 through February 28, no person shall possess more than 15 black sea bass in, or harvested from, the...

  9. 50 CFR 648.145 - Black sea bass possession limit.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 50 Wildlife and Fisheries 12 2013-10-01 2013-10-01 false Black sea bass possession limit. 648.145... Measures for the Black Sea Bass Fishery § 648.145 Black sea bass possession limit. (a) During the recreational fishing season specified at § 648.146, no person shall possess more than 20 black sea bass in,...

  10. 50 CFR 648.145 - Black sea bass possession limit.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 50 Wildlife and Fisheries 12 2014-10-01 2014-10-01 false Black sea bass possession limit. 648.145... Measures for the Black Sea Bass Fishery § 648.145 Black sea bass possession limit. (a) During the recreational fishing season specified at § 648.146, no person shall possess more than 15 black sea bass in,...

  11. The contextuality of the possessed values

    NASA Astrophysics Data System (ADS)

    Kim, Myung Suk; Jo, Sang Gyu; Choi, Sang Don; Kim, Jung Ho; Moo Jeon, Hun; Kim, Ho Il

    1999-04-01

    The mathematical structure of quantum mechanics is determined by two algorithms: quantization algorithm (QUAN) which says that an observed value of an observable Q is one of the eigenvalues q of the corresponding physical operator icons/Journals/Common/hatQ" ALT="hatQ" ALIGN="TOP"/>, and the statistical algorithm (STAT) which says that the probability for the observed physical quantity vm(Q) to be q is |icons/Journals/Common/langle" ALT="langle" ALIGN="TOP"/> q|icons/Journals/Common/psi" ALT="psi" ALIGN="TOP"/>icons/Journals/Common/rangle" ALT="rangle" ALIGN="TOP"/>|2. From these two algorithms we can derive the principle of statistical function composition (SFC), Prob(vm(f(Q)) = r|icons/Journals/Common/psi" ALT="psi" ALIGN="TOP"/>) = Prob(vm(Q) = f-1(r)|icons/Journals/Common/psi" ALT="psi" ALIGN="TOP"/>). With the assumption of the principle of function composition (FC) that the algebraic relations between observables of a given physical system are identical with the algebraic relations between the values possessed by the system before the observation is made, it could be explained why the result of measurement obeys SFC. According to Bell-KS theorem, FC is not compatible with quantum theory. We analyse, in this paper, the incompatibility of FC with quantum theory in detail, focusing on the analysis of Redhead (1989 Incompleteness, Nonlocality and Realism (Oxford: Oxford University Press)). Redhead assumes the faithful measurement principle (FM) in order to construct FC. However, we show, that FC can be derived without the help of FM.

  12. Metabolic Syndrome

    MedlinePLUS

    ... page from the NHLBI on Twitter. What Is Metabolic Syndrome? Metabolic syndrome is the name for a group of risk ... three metabolic risk factors to be diagnosed with metabolic syndrome. A large waistline. This also is called abdominal ...

  13. Metabolic syndrome

    MedlinePLUS

    Metabolic syndrome is a name for a group of risk factors that occur together and increase the chance ... Metabolic syndrome is becoming very common in the United States. Doctors are not sure whether the syndrome is ...

  14. Gardner Syndrome

    MedlinePLUS

    ... Gardner syndrome? Gardner syndrome is a subtype of familial adenomatous polyposis (FAP or classic FAP) , which usually causes benign, ... is linked to Gardner syndrome; APC stands for adenomatous polyposis coli. A mutation, meaning an alteration in the APC ...

  15. Down Syndrome

    MedlinePLUS

    ... NICHD Research Information Clinical Trials Resources and Publications Down Syndrome: Condition Information Skip sharing on social media links Share this: Page Content What is Down syndrome? Down syndrome describes a set of cognitive and ...

  16. Down Syndrome

    MedlinePLUS

    ... Digestive System How the Body Works Main Page Down Syndrome KidsHealth > Kids > Health Problems > Birth Defects & Genetic Problems > ... skills. Continue Do a Lot of People Have Down Syndrome? Down syndrome is not contagious , so you can' ...

  17. Cushing's Syndrome

    MedlinePLUS

    ... being done? Clinical Trials Organizations What is Cushing's Syndrome? Cushing's syndrome, also called hypercortisolism , is a rare endocrine ... and cure the disorder. NIH Patient Recruitment for Cushing's Syndrome Clinical Trials At NIH Clinical Center Throughout the ...

  18. Brown Syndrome

    MedlinePLUS

    ... Does Brown syndrome cause eye problems besides abnormal eye movements? Some children with Brown syndrome have poor binocular ... In the congenital form of Brown syndrome, the eye movement problem is usually constant and unlikely to resolve ...

  19. Dravet Syndrome

    MedlinePLUS

    ... NINDS Dravet Syndrome Information Page Synonym(s): Severe Myoclonic Epilepsy of Infancy (SMEI) Table of Contents (click to ... Dravet Syndrome? Dravet syndrome, also called severe myoclonic epilepsy of infancy (SMEI), is a severe form of ...

  20. A 'second life' agenda. Psychiatric research issues raised by protease inhibitor treatments for people with the human immunodeficiency virus or the acquired immunodeficiency syndrome.

    PubMed

    Rabkin, J G; Ferrando, S

    1997-11-01

    Seldom in the history of medicine has an entire generation of patients with an incurable, progressive, and ultimately fatal disease suddenly been offered the prospect of extended survival and even, perhaps, a "second life." The relatively simultaneous appearance of 2 major treatment developments has created profound changes in therapeutic options and outlook. The first development is an assay of serum levels of human immunodeficiency virus viral copies, providing a critical tool for clinical decision making. The second is the marketing between December 1995 and April 1997 of 4 human immunodeficiency virus protease inhibitors that, combined with previously available antiviral medications, achieve a new level of efficacy. With the advent of these changes come multiple psychiatric research and policy issues. These include the development of strategies to establish and maintain medication adherence. This is a critical task, given the complexity of combination therapy regimens and the rapid onset of viral resistance to protease inhibitors within days to weeks of missed or suboptimal dosing. The psychological issues to be studied include the process of restructuring lives and expectations in the event of clinical benefit or managing the distress associated with clinical failure. Other research questions include the effects of restored health on the appraisal of human immunodeficiency virus risk behaviors, assessment of effect of neurocognitive functioning, and unanswered questions about psychotropic or protease inhibitor drug interactions due to their shared metabolic pathways. Behavioral scientists can inform provision of care to patients who may be considered difficult to treat, such as those with severe and persistent mental illness or active substance abuse or the homeless. This includes the provision of empirical data regarding individual and situational characteristics that are likely to promote or impede adherence, as well as innovative provision systems. Psychiatry can make notable contributions during this turning point in human immunodeficiency virus therapeutics and research. PMID:9366663

  1. Variation in Fetal Outcome, Viral Load and ORF5 Sequence Mutations in a Large Scale Study of Phenotypic Responses to Late Gestation Exposure to Type 2 Porcine Reproductive and Respiratory Syndrome Virus

    PubMed Central

    Ladinig, Andrea; Wilkinson, Jamie; Ashley, Carolyn; Detmer, Susan E.; Lunney, Joan K.; Plastow, Graham; Harding, John C. S.

    2014-01-01

    In spite of extensive research, the mechanisms of reproductive disease associated with Porcine Reproductive and Respiratory Syndrome virus (PRRSv) are still poorly understood. The objectives of this large scale study were to evaluate associations between viral load and fetal preservation, determine the impact of type 2 PRRSv on fetal weights, and investigate changes in ORF5 PRRSv genome in dams and fetuses during a 21-day period following challenge. At gestation day 85 (±1), 114 gilts were experimentally infected with type 2 PRRSv, while 19 gilts served as reference controls. At necropsy, fetuses were categorized according to their preservation status and tissue samples were collected. PRRSv RNA concentrations were measured in gilt serum collected on days 0, 2, 6, and 21 post-infection, as well as in gilt and fetal tissues collected at termination. Fetal mortality was 41±22.8% in PRRS infected litters. Dead fetuses appeared to cluster in some litters but appeared solitary or random in others. Nine percent of surviving piglets were meconium-stained. PRRSv RNA concentration in fetal thymus, fetal serum and endometrium differed significantly across preservation category and was greatest in tissues of meconium-stained fetuses. This, together with the virtual absence of meconium staining in non-infected litters indicates it is an early pathological condition of reproductive PRRS. Viral load in fetal thymus and in fetal serum was positively associated with viral load in endometrium, suggesting the virus exploits dynamic linkages between individual maternal-fetal compartments. Point mutations in ORF5 sequences from gilts and fetuses were randomly located in 20 positions in ORF5, but neither nucleotide nor amino acid substitutions were associated with fetal preservation. PRRSv infection decreased the weights of viable fetuses by approximately 17%. The considerable variation in gilt and fetal outcomes provides tremendous opportunity for more detailed investigations of potential mechanisms and single nucleotide polymorphisms associated with fetal death. PMID:24756023

  2. Hereditary orotic aciduria, Lesch-Nyhan syndrome, and xeroderma pigmentosum probed by herpes simplex virus: /sup 125/I-iododeoxycytidine incorporation as an assay for viral growth. [Human fibroblasts

    SciTech Connect

    Campisi, J.; Hafner, J.; Boorstein, R.; Pardee, A.B.

    1983-01-01

    /sup 125/I-Iododeoxycytidine (/sup 125/IdC) incorporation into acid-insoluble material was a sensitive, rapid, and quantitative assay for the growth of herpes simplex virus type 1 (HSV-1) in human fibroblasts. Cellular utilization of the isotope was 10 to 25% of the incorporation by infected cells and could be 80% inhibited by tetrahydrouridine (THU). Viral utilization was inhibited by acycloguanosine, thioguanine (TG), and cytosine arabinoside. Isotope was incorporated equally well by growing or quiescent infected cells. HSV-1 was used to probe the metabolic capabilities of three mutant human fibroblast strains. /sup 125/IdC incorporation quantitatively measured the ability of the virus to grow in these cells. Viral /sup 125/IdC incorporation was sensitive to TG in normal fibroblasts but showed a 8- to 10-fold greater resistance to TG in fibroblasts derived from patients with Lesch-Nyhan syndrome (LN). Similarly, the growth of ultraviolet irradiated HSV-1 in normal fibroblasts was 5-fold greater than in fibroblasts derived from patients with xeroderma pigmentosum. In fibroblasts derived from patients with hereditary orotic aciduria, viral /sup 125/IdC incorporation was sensitive to adenosine (AD) at concentrations which were slightly stimulatory in normal fibroblasts. This was a 2-fold difference in AD sensitivity, which the radioassay reliably and quantitatively documented. HSV-1 infected cells could be individually identified by their incorporated /sup 125/IdC; such cells had blackened nuclei in autoradiograms prepared 12 hr after infection. Normal cells infected in the presence of TG had many fewer labeled nuclei than LN cells similarly infected in the presence of the drug. (JMT)

  3. Diagnosis of the Lelystad strain of Porcine reproductive and respiratory syndrome virus infection in individually housed pigs: comparison between serum and oral fluid samples for viral nucleic acid and antibody detection.

    PubMed

    Decorte, Inge; Van Campe, Willem; Mostin, Laurent; Cay, Ann Brigitte; De Regge, Nick

    2015-01-01

    There has been a developing interest in the use of oral fluid for the diagnosis of different pathogens such as Porcine reproductive and respiratory syndrome virus (PRRSV). PRRSV and PRRSV-specific antibodies have been shown to be present in oral fluid samples, but the correlation between diagnostic results in oral fluid and serum samples has been insufficiently addressed. Studies investigating this correlation focused on boars older than 6 months and type 2 strains, but it is known that the outcome of a PRRSV infection is age and strain dependent. To address this gap, the current study reports on the detection of PRRSV and PRRSV-specific antibodies in serum and oral fluid samples collected over a 6-week period after an experimental infection of 8-week-old individually housed pigs with Lelystad virus, the type 1 prototype strain. Quantitative reverse transcription polymerase chain reaction analysis showed that significantly more serum samples were PRRSV RNA-positive than oral fluid until 5 days postinfection (dpi). Between 7 and 21 dpi, PRRSV RNA detection was similar in both samples but higher detection rates in oral fluid were found from 28 dpi. Compared with existing literature, this highlights that detection rates at particular time points postinfection might vary in function of strain virulence and animal age and provides useful information for the interpretation of pen-based oral fluid results. An excellent agreement between the oral fluid and serum enzyme-linked immunosorbent assay results was observed at every time point, further supporting the usefulness of oral fluid as a diagnostic sample for antibody detection. PMID:25525137

  4. Pathogenesis of human immunodeficiency virus infection and prospects for control.

    PubMed Central

    Ho, D. D.; Kaplan, J. C.

    1987-01-01

    In just six years after the initial description of the acquired immunodeficiency syndrome, much has been learned about the etiologic agent, the human immunodeficiency virus. The pathogenic mechanisms utilized by this virus to infect selectively and persistently T4+ lymphocytes and monocyte/macrophages, leading to immunodeficiency and neurologic dysfunction, are slowly becoming clear. Better understanding of the pathogenesis of human immunodeficiency virus infection is essential for the rational design of therapeutic and preventive strategies to combat this deadly virus. PMID:3324508

  5. Role of CD4+ and CD8+ T-Cell Responses against JC Virus in the Outcome of Patients with Progressive Multifocal Leukoencephalopathy (PML) and PML with Immune Reconstitution Inflammatory Syndrome ?

    PubMed Central

    Gheuens, Sarah; Bord, Evelyn; Kesari, Santosh; Simpson, David M.; Gandhi, Rajesh T.; Clifford, David B.; Berger, Joseph R.; Ngo, Long; Koralnik, Igor J.

    2011-01-01

    Progressive multifocal leukoencephalopathy (PML) is a severe demyelinating disease of the brain caused by JC virus (JCV). To assess the role of CD4+ and CD8+ T-cells against JCV in the clinical outcome of PML and PML in the setting of immune reconstitution inflammatory syndrome (IRIS), we tested gamma interferon (IFN-?) response by enzyme-linked immunosorbent spot (ELISpot) and intracellular cytokine staining (ICS) in 117 subjects, including 66 PML patients with different clinical outcomes. Both assays were concordant and demonstrated that the cellular immune response against JCV is associated with better clinical outcome. PML survivors had an early CD8+ T-cell response more frequently than PML progressors (100% versus 27.3%; P = 0.001), while only a trend was observed for the early CD4+ T-cell response between these two groups (80% versus 45.5%; P = 0.18). Although IRIS itself was more frequent in the PML survivor group, there was no difference in IFN-?-producing CD4+ and CD8+ T-cells between IRIS and non-IRIS PML patients, suggesting that T-cells expressing other cytokines likely have a role in the immunopathogenesis of IRIS. ELISpot and ICS assays are useful prognostic markers of PML evolution and may help in the clinical management of these patients. PMID:21543472

  6. Evaluation of the immunogenicity of a transgenic tobacco plant expressing the recombinant fusion protein of GP5 of porcine reproductive and respiratory syndrome virus and B subunit of Escherichia coli heat-labile enterotoxin in pigs.

    PubMed

    Chia, Min-Yuan; Hsiao, Shih-Hsuan; Chan, Hui-Ting; Do, Yi-Yin; Huang, Pung-Ling; Chang, Hui-Wen; Tsai, Yi-Chieh; Lin, Chun-Ming; Pang, Victor Fei; Jeng, Chian-Ren

    2011-04-15

    Escherichia coli heat-labile enterotoxin B subunit (LTB) can be used as an adjuvant for co-administered antigens. Our previous study showed that the expression of neutralizing epitope GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) in transgenic tobacco plant (GP5-T) could induce PRRSV-specific immune responses in pigs. A transgenic tobacco plant co-expressing LTB and PRRSV GP5 as a fusion protein (LTB-GP5-T) was further constructed and its immunogenicity was evaluated. Pigs were given orally three consecutive doses of equal concentration of recombinant GP5 protein expressed in leaves of LTB-GP5-T or GP5-T at a 2-week interval and challenged with PRRSV at 7 weeks post-initial immunization. Pigs receiving LTB-GP5-T or GP5-T developed PRRSV-specific antibody- and cell-mediated immunity and showed significantly lower viremia and tissue viral load and milder lung lesions than wild type tobacco plant (W-T). The LTB-GP5-T-treated group had relatively higher immune responses than the GP5-T-treated group, although the differences were not statistically significant. PMID:21277027

  7. Evaluation of monoclonal antibody-based immunohistochemistry for the detection of European and North American Porcine reproductive and respiratory syndrome virus and a comparison with in situ hybridization and reverse transcription polymerase chain reaction.

    PubMed

    Han, Kiwon; Seo, Hwi Won; Oh, Yeonsu; Kang, Ikjae; Park, Changhoon; Kang, Sang Hoon; Kim, Sung-Hoon; Lee, Bog-Hieu; Kwon, Byungjoon; Chae, Chanhee

    2012-07-01

    The objective of the present study was to compare the ability of 2 monoclonal antibodies (mAbs; SDOW17 and SR30) to detect types 1 and 2 Porcine reproductive and respiratory syndrome virus (PRRSV) in formalin-fixed, paraffin-embedded (FFPE) lung tissues by immunohistochemistry (IHC) and to compare the immunohistochemical results with in situ hybridization (ISH) and reverse transcription nested polymerase chain reaction (RT-nPCR) detection techniques. Lungs from 30 experimentally infected pigs (15 pigs with each genotype of PRRSV) and 20 naturally infected pigs (10 pigs with each genotype of PRRSV) with types 1 and 2 PRRSV, respectively, were used for the IHC, ISH, and RT-nPCR analyses. The SR30 mAb-based IHC detected significantly more type 1 PRRSV-positive cells in the accessory and caudal lobes from the experimentally infected pigs at 7 (P = 0.025) and 14 (P = 0.018) days postinoculation, respectively, compared to the SDOW17 mAb-based IHC. The results demonstrated that SR30 mAb-based IHC is useful for detecting both types 1 and 2 PRRSV antigen in FFPE lung tissues. PMID:22649158

  8. [Syndromes 12. Turner syndrome].

    PubMed

    Verdonck, A; van Erum, R

    1999-07-01

    Turner syndrome is one of the most common chromosomal disorders. The incidence is about 1 on 2,500 till 1 on 10,000 living female young births. Short stature is the most common finding in patients with Turner syndrome. Besides short stature and gonadal dysgenesis, typical craniofacial and dental features are also present. Disturbance of the enchondral ossification results in abnormal craniofacial morphology. Oestrogen medication, to induce their puberty, and recombinant human growth therapy, to improve final height of these patients, are the most common treatment possibilities. It is the intention of this short paper to inform the dentist/orthodontist about the general aspects of the Turner syndrome. This information can be used in their treatment plan. PMID:11930372

  9. Foodborne viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Testing for human pathogenic viruses in foods represents a formidable task requiring the extraction, concentration, and assay of a host of viruses from a wide range of food matrices. The enteric viruses, particularly genogroup I and II (GI and GII) noroviruses and hepatitis A virus, are the princip...

  10. Systems analysis of West Nile virus infection

    PubMed Central

    Suthar, Mehul S.; Pulendran, Bali

    2014-01-01

    Emerging and re-emerging mosquito-borne viruses continue to pose a significant threat to human health throughout the world. Over the past decade, West Nile virus (WNV), Dengue virus (DENV), and Chikungunya virus (CHIKV), have caused annual epidemics of virus-induced encephalitis, hemorrhagic fever\\shock syndromes, and arthritis, respectively. Currently, no specific antiviral therapies or vaccines exist for use in humans to combat or prevent these viral infections. Thus, there is a pressing need to define the virus-host interactions that govern immunity and infection outcome. Recent technological breakthroughs in ‘omics’ resources and high-throughput based assays are beginning to accelerate antiviral drug discovery and improve on current strategies for vaccine design. In this review, we highlight studies with WNV and discuss how traditional and systems based approaches are being used to rapidly identify novel host targets for therapeutic intervention and develop a deeper conceptual understanding of the host response to virus infection. PMID:24851811

  11. Ramsay Hunt syndrome.

    PubMed

    Sweeney, C J; Gilden, D H

    2001-08-01

    The strict definition of the Ramsay Hunt syndrome is peripheral facial nerve palsy accompanied by an erythematous vesicular rash on the ear (zoster oticus) or in the mouth. J Ramsay Hunt, who described various clinical presentations of facial paralysis and rash, also recognised other frequent symptoms and signs such as tinnitus, hearing loss, nausea, vomiting, vertigo, and nystagmus. He explained these eighth nerve features by the close proximity of the geniculate ganglion to the vestibulocochlear nerve within the bony facial canal. Hunt's analysis of clinical variations of the syndrome now bearing his name led to his recognition of the general somatic sensory function of the facial nerve and his defining of the geniculate zone of the ear. It is now known that varicella zoster virus (VZV) causes Ramsay Hunt syndrome. Compared with Bell's palsy (facial paralysis without rash), patients with Ramsay Hunt syndrome often have more severe paralysis at onset and are less likely to recover completely. Studies suggest that treatment with prednisone and acyclovir may improve outcome, although a prospective randomised treatment trial remains to be undertaken. In the only prospective study of patients with Ramsay Hunt syndrome, 14% developed vesicles after the onset of facial weakness. Thus, Ramsay Hunt syndrome may initially be indistinguishable from Bell's palsy. Further, Bell's palsy is significantly associated with herpes simplex virus (HSV) infection. In the light of the known safety and effectiveness of antiviral drugs against VZV or HSV, consideration should be given to early treatment of all patients with Ramsay Hunt syndrome or Bell's palsy with a 7-10 day course of famciclovir (500 mg, three times daily) or acyclovir (800 mg, five times daily), as well as oral prednisone (60 mg daily for 3-5 days). Finally, some patients develop peripheral facial paralysis without ear or mouth rash, associated with either a fourfold rise in antibody to VZV or the presence of VZV DNA in auricular skin, blood mononuclear cells, middle ear fluid, or saliva. This indicates that a proportion of patients with "Bell's palsy" have Ramsay Hunt syndrome zoster sine herpete. Treatment of these patients with acyclovir and prednisone within 7 days of onset has been shown to improve the outcome of recovery from facial palsy. PMID:11459884

  12. Hepatitis C virus syndrome: A constellation of organ- and non-organ specific autoimmune disorders, B-cell non-Hodgkin’s lymphoma, and cancer

    PubMed Central

    Ferri, Clodoveo; Sebastiani, Marco; Giuggioli, Dilia; Colaci, Michele; Fallahi, Poupak; Piluso, Alessia; Antonelli, Alessandro; Zignego, Anna Linda

    2015-01-01

    The clinical course of chronic hepatitis C virus (HCV) infection is characterized by possible development of both liver and extrahepatic disorders. The tropism of HCV for the lymphoid tissue is responsible for several immune-mediated disorders; a poly-oligoclonal B-lymphocyte expansion, commonly observed in a high proportion of patients with HCV infection, are responsible for the production of different autoantibodies and immune-complexes, such as mixed cryoglobulins. These serological alterations may characterize a variety of autoimmune or neoplastic diseases. Cryoglobulinemic vasculitis due to small-vessel deposition of circulating mixed cryoglobulins is the prototype of HCV-driven immune-mediated and lymphoproliferative disorders; interestingly, in some cases the disease may evolve to frank malignant lymphoma. In addition, HCV shows an oncogenic potential as suggested by several clinico-epidemiological and laboratory studies; in addition to hepatocellular carcinoma that represents the most frequent HCV-related malignancy, a causative role of HCV has been largely demonstrated in a significant percentage of patients with isolated B-cells non-Hodgkin’s lymphomas. The same virus may be also involved in the pathogenesis of papillary thyroid cancer, a rare neoplastic condition that may complicate HCV-related thyroid involvement. Patients with HCV infection are frequently asymptomatic or may develop only hepatic alteration, while a limited but clinically relevant number can develop one or more autoimmune and/or neoplastic disorders. Given the large variability of their prevalence among patients’ populations from different countries, it is possible to hypothesize a potential role of other co-factors, i.e., genetic and/or environmental, in the pathogenesis of HCV-related extra-hepatic diseases. PMID:25848462

  13. 50 CFR 622.408 - Bag/possession limits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE CARIBBEAN, GULF OF MEXICO, AND SOUTH ATLANTIC Spiny Lobster Fishery of the Gulf of Mexico and South Atlantic § 622.408 Bag/possession limits. (a) EEZ off the... or possession limit of spiny lobster in or from the EEZ off Florida and off the Gulf states,...

  14. 50 CFR 648.40 - Prohibition on possession.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Atlantic Salmon § 648.40 Prohibition on possession. (a) Incidental catch. All Atlantic salmon caught... maximum probability of survival. (b) Presumption. The possession of Atlantic salmon is prima facie evidence that such Atlantic salmon were taken in violation of this regulation. Evidence that such fish...

  15. 50 CFR 648.40 - Prohibition on possession.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Atlantic Salmon § 648.40 Prohibition on possession. (a) Incidental catch. All Atlantic salmon caught... maximum probability of survival. (b) Presumption. The possession of Atlantic salmon is prima facie evidence that such Atlantic salmon were taken in violation of this regulation. Evidence that such fish...

  16. 50 CFR 648.40 - Prohibition on possession.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Atlantic Salmon § 648.40 Prohibition on possession. (a) Incidental catch. All Atlantic salmon caught... maximum probability of survival. (b) Presumption. The possession of Atlantic salmon is prima facie evidence that such Atlantic salmon were taken in violation of this regulation. Evidence that such fish...

  17. 50 CFR 648.40 - Prohibition on possession.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Atlantic Salmon § 648.40 Prohibition on possession. (a) Incidental catch. All Atlantic salmon caught... maximum probability of survival. (b) Presumption. The possession of Atlantic salmon is prima facie evidence that such Atlantic salmon were taken in violation of this regulation. Evidence that such fish...

  18. 50 CFR 648.40 - Prohibition on possession.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Atlantic Salmon § 648.40 Prohibition on possession. (a) Incidental catch. All Atlantic salmon caught... maximum probability of survival. (b) Presumption. The possession of Atlantic salmon is prima facie evidence that such Atlantic salmon were taken in violation of this regulation. Evidence that such fish...

  19. The Relationship between Social Capital and Weapon Possession on Campus

    ERIC Educational Resources Information Center

    Messer, Rachel H.; Bradley, Kristopher I.; Calvi, Jessica L.; Kennison, Shelia M.

    2012-01-01

    The present research focused on the problem of how college officials might be able to predict weapon possession on college campuses. We hypothesized that measures of social capital (i.e., trust and participation in society) may be useful in identifying individuals who are likely to possess weapons on campuses. Prior research has shown that those…

  20. 50 CFR 622.408 - Bag/possession limits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...OF MEXICO, AND SOUTH ATLANTIC Spiny Lobster Fishery of the Gulf of Mexico and South...daily bag or possession limit for spiny lobster in or from the EEZ off the southern...daily bag or possession limit of spiny lobster in or from the EEZ off Florida and...

  1. 50 CFR 622.408 - Bag/possession limits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...OF MEXICO, AND SOUTH ATLANTIC Spiny Lobster Fishery of the Gulf of Mexico and South...daily bag or possession limit for spiny lobster in or from the EEZ off the southern...daily bag or possession limit of spiny lobster in or from the EEZ off Florida and...

  2. 50 CFR 622.408 - Bag/possession limits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... ADMINISTRATION, DEPARTMENT OF COMMERCE FISHERIES OF THE CARIBBEAN, GULF OF MEXICO, AND SOUTH ATLANTIC Spiny Lobster Fishery of the Gulf of Mexico and South Atlantic § 622.408 Bag/possession limits. (a) EEZ off the... or possession limit of spiny lobster in or from the EEZ off Florida and off the Gulf states,...

  3. 9. Photocopy of 1845 manuscript (original in the possession of ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    9. Photocopy of 1845 manuscript (original in the possession of the Metropolitan Museum of Art, New York City, N.Y.; photocopy in the possession of the Onondaga Historical Association) ENTRY IN A. J. DAVIS' (ARCHITECT) ACCOUNT BOOK, SHOWING SKETCH OF WEST ELEVATION AND FIRST LEVEL PLAN, AND SCHEDULE OF TEN DRAWINGS - Sedgewick House, 742 James Street, Syracuse, Onondaga County, NY

  4. 50 CFR 648.235 - Possession and landing restrictions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...governing the harvest, possession, landing, purchase, and sale of shark fins are found at part 600, subpart N, of this chapter...governing the harvest, possession, landing, purchase, and sale of shark fins are found at part 600, subpart N, of this...

  5. Whose? L2-English Speakers' Possessive Pronoun Gender Errors

    ERIC Educational Resources Information Center

    Anton-Mendez, Ines

    2011-01-01

    This article reports the results of an experiment on production of "his/her" in English as a second language (L2) by proficient native speakers of Italian, Spanish, and Dutch. In Dutch and English, 3rd person singular possessive pronouns agree in gender with their antecedents, in Italian and Spanish possessives in general agree with the noun they…

  6. 45 CFR 670.6 - Prior possession exception.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 45 Public Welfare 3 2014-10-01 2014-10-01 false Prior possession exception. 670.6 Section 670.6 Public Welfare Regulations Relating to Public Welfare (Continued) NATIONAL SCIENCE FOUNDATION CONSERVATION OF ANTARCTIC ANIMALS AND PLANTS Prohibited Acts, Exceptions § 670.6 Prior possession exception. (a) Exception. Section 670.4 shall not apply...

  7. 2012 Boise State University 1 Possession of Firearms/Weapons

    E-print Network

    Barrash, Warren

    © 2012 Boise State University 1 Possession of Firearms/Weapons on University Owned or Controlled for the regulation of the possession of firearms and other weapons on university owned and controlled premises · Scope ­ applies to all firearms and other weapons on university owned or controlled premises

  8. 50 CFR 635.30 - Possession at sea and landing.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...or operate a fishing vessel that possesses an Atlantic tuna in the Atlantic Ocean or that lands an Atlantic tuna in an Atlantic...fishing vessel that possesses a swordfish in the Atlantic Ocean or lands a swordfish in an Atlantic coastal...

  9. 50 CFR 635.30 - Possession at sea and landing.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...or operate a fishing vessel that possesses an Atlantic tuna in the Atlantic Ocean or that lands an Atlantic tuna in an Atlantic...fishing vessel that possesses a swordfish in the Atlantic Ocean or lands a swordfish in an Atlantic coastal...

  10. 50 CFR 635.30 - Possession at sea and landing.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...or operate a fishing vessel that possesses an Atlantic tuna in the Atlantic Ocean or that lands an Atlantic tuna in an Atlantic...fishing vessel that possesses a swordfish in the Atlantic Ocean or lands a swordfish in an Atlantic coastal...

  11. 50 CFR 635.30 - Possession at sea and landing.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...or operate a fishing vessel that possesses an Atlantic tuna in the Atlantic Ocean or that lands an Atlantic tuna in an Atlantic...fishing vessel that possesses a swordfish in the Atlantic Ocean or lands a swordfish in an Atlantic coastal...

  12. Pre-Posed Possessive Constructions in Russian and Polish

    ERIC Educational Resources Information Center

    Houle, Erik Richard

    2013-01-01

    In Contemporary Standard Russian (CSR) and Contemporary Standard Polish (CSP) nominal possession is conveyed by means of the adnominal genitive. In this construction the dependent follows the noun it modifies and is marked morphologically for possession in the genitive case. The head noun is marked morphologically for any one of the six…

  13. Possession experiences in dissociative identity disorder: a preliminary study.

    PubMed

    Ross, Colin A

    2011-01-01

    Dissociative trance disorder, which includes possession experiences, was introduced as a provisional diagnosis requiring further study in the Diagnostic and Statistical Manual of Mental Disorders (4th ed.). Consideration is now being given to including possession experiences within dissociative identity disorder (DID) in the Diagnostic and Statistical Manual of Mental Disorders (5th ed.), which is due to be published in 2013. In order to provide empirical data relevant to the relationship between DID and possession states, I analyzed data on the prevalence of trance, possession states, sleepwalking, and paranormal experiences in 3 large samples: patients with DID from North America; psychiatric outpatients from Shanghai, China; and a general population sample from Winnipeg, Canada. Trance, sleepwalking, paranormal, and possession experiences were much more common in the DID patients than in the 2 comparison samples. The study is preliminary and exploratory in nature because the samples were not matched in any way. PMID:21667381

  14. Structure-Activity Relationships in the Binding of Chemically Derivatized CD4 to gp120 from Human Immunodeficiency Virus

    E-print Network

    Hendrickson, Wayne A.

    immunodeficiency virus 1 (HIV-1a) is the primary cause of acquired immunodeficiency syndrome (AIDS).1,2 Among 20 Immunodeficiency Virus Hui Xie,#,3 Danny Ng,§,| Sergey N. Savinov,§, Barna Dey, Peter D. Kwong, Richard Wyatt, Amos

  15. A Novel Virus Causes Scale Drop Disease in Lates calcarifer.

    PubMed

    de Groof, Ad; Guelen, Lars; Deijs, Martin; van der Wal, Yorick; Miyata, Masato; Ng, Kah Sing; van Grinsven, Lotte; Simmelink, Bartjan; Biermann, Yvonne; Grisez, Luc; van Lent, Jan; de Ronde, Anthony; Chang, Siow Foong; Schrier, Carla; van der Hoek, Lia

    2015-08-01

    From 1992 onwards, outbreaks of a previously unknown illness have been reported in Asian seabass (Lates calcarifer) kept in maricultures in Southeast Asia. The most striking symptom of this emerging disease is the loss of scales. It was referred to as scale drop syndrome, but the etiology remained enigmatic. By using a next-generation virus discovery technique, VIDISCA-454, sequences of an unknown virus were detected in serum of diseased fish. The near complete genome sequence of the virus was determined, which shows a unique genome organization, and low levels of identity to known members of the Iridoviridae. Based on homology of a series of putatively encoded proteins, the virus is a novel member of the Megalocytivirus genus of the Iridoviridae family. The virus was isolated and propagated in cell culture, where it caused a cytopathogenic effect in infected Asian seabass kidney and brain cells. Electron microscopy revealed icosahedral virions of about 140 nm, characteristic for the Iridoviridae. In vitro cultured virus induced scale drop syndrome in Asian seabass in vivo and the virus could be reisolated from these infected fish. These findings show that the virus is the causative agent for the scale drop syndrome, as each of Koch's postulates is fulfilled. We have named the virus Scale Drop Disease Virus. Vaccines prepared from BEI- and formalin inactivated virus, as well as from E. coli produced major capsid protein provide efficacious protection against scale drop disease. PMID:26252390

  16. A Novel Virus Causes Scale Drop Disease in Lates calcarifer

    PubMed Central

    de Groof, Ad; Guelen, Lars; Deijs, Martin; van der Wal, Yorick; Miyata, Masato; Ng, Kah Sing; van Grinsven, Lotte; Simmelink, Bartjan; Biermann, Yvonne; Grisez, Luc; van Lent, Jan; de Ronde, Anthony; Chang, Siow Foong; Schrier, Carla; van der Hoek, Lia

    2015-01-01

    From 1992 onwards, outbreaks of a previously unknown illness have been reported in Asian seabass (Lates calcarifer) kept in maricultures in Southeast Asia. The most striking symptom of this emerging disease is the loss of scales. It was referred to as scale drop syndrome, but the etiology remained enigmatic. By using a next-generation virus discovery technique, VIDISCA-454, sequences of an unknown virus were detected in serum of diseased fish. The near complete genome sequence of the virus was determined, which shows a unique genome organization, and low levels of identity to known members of the Iridoviridae. Based on homology of a series of putatively encoded proteins, the virus is a novel member of the Megalocytivirus genus of the Iridoviridae family. The virus was isolated and propagated in cell culture, where it caused a cytopathogenic effect in infected Asian seabass kidney and brain cells. Electron microscopy revealed icosahedral virions of about 140 nm, characteristic for the Iridoviridae. In vitro cultured virus induced scale drop syndrome in Asian seabass in vivo and the virus could be reisolated from these infected fish. These findings show that the virus is the causative agent for the scale drop syndrome, as each of Koch’s postulates is fulfilled. We have named the virus Scale Drop Disease Virus. Vaccines prepared from BEI- and formalin inactivated virus, as well as from E. coli produced major capsid protein provide efficacious protection against scale drop disease. PMID:26252390

  17. Craniofacial Syndrome Descriptions

    MedlinePLUS

    ... brought about by birth defect, disease or trauma. Apert syndrome Carpenter syndrome Carpenter Syndrome belongs to a group ... FAQs CCAkids Blog CCA Web Store Cher Syndromes • Apert syndrome • Carpenter syndrome • Cleft lip and/or palate • Craniosynostosis • ...

  18. Analysis of ORF5 and Full-Length Genome Sequences of Porcine Reproductive and Respiratory Syndrome Virus Isolates of Genotypes 1 and 2 Retrieved Worldwide Provides Evidence that Recombination Is a Common Phenomenon and May Produce Mosaic Isolates

    PubMed Central

    Martín-Valls, G. E.; Kvisgaard, L. K.; Tello, M.; Darwich, L.; Cortey, M.; Burgara-Estrella, A. J.; Hernández, J.; Larsen, L. E.

    2014-01-01

    ABSTRACT Recombination is currently recognized as a factor for high genetic diversity, but the frequency of such recombination events and the genome segments involved are not well known. In the present study, we initially focused on the detection of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) isolates by examining previously published data sets of ORF5 sequences (genotypes 1 and 2) obtained worldwide. We then examined full-length genome sequences in order to determine potential recombination breakpoints along the viral genome. For ORF5, 11 sets of genotype 1 sequences from different geographical areas, including 2 Asian, 1 American, and 7 European regions, and three sets of genotype 2, including sets from China, Mexico, and the United States, were analyzed separately. Potential recombination breakpoints were detected in 10/11 genotype 1 sets, including 9 cases in which the clustering of at least one isolate was different before and after the breakpoints. In genotype 2, potential breakpoints and different tree clustering of at least one strain before and after the breakpoint were observed in 2 out of 3 sets. The results indicated that most of the ORF5 data sets contained at least one recombinant sequence. When the full-length genome sequences were examined, both genotype 1 and 2 sets presented breakpoints (10 and 9, respectively), resulting in significantly different topologies before and after the breakpoints. Mosaic genomes were detected in genotype 1 sequences. These results may have significant implications for the understanding of the molecular epidemiology of PRRSV. IMPORTANCE PRRSV is one of the most important viruses affecting swine production worldwide, causing big economic losses and sanitary problems. One of the key questions on PRRSV arises from its genetic diversity, which is thought to have a direct impact on immunobiology, epidemiology, diagnosis, and vaccine efficacy. One of the causes of this genetic diversity is recombination among strains. This study provides evidence that recombinant PRRSV isolates are common in most of the countries with significant swine production, especially PRRSV genotype 1. This observation has implications in the proper characterization of PRRSV strains, in the future development of phylogenetic studies, and in the development of new PRRSV control strategies. Moreover, the present paper emphasizes the need for a deeper understanding of the mechanisms and circumstances involved in the generation of genetic diversity of PRRSV. PMID:24371078

  19. Usher Syndrome

    MedlinePLUS

    Usher syndrome is an inherited disease that causes serious hearing loss and retinitis pigmentosa, an eye disorder ... hearing and vision. There are three types of Usher syndrome: People with type I are deaf from ...

  20. Hunter syndrome

    MedlinePLUS

    ... form: Mild to no mental deficiency Both forms: Carpal tunnel syndrome Coarse features of the face Deafness (gets worse ... Airway obstruction Carpal tunnel syndrome Hearing loss that gets worse ... to complete daily living activities Joint stiffness that ...

  1. Klinefelter Syndrome

    MedlinePLUS

    ... Information Clinical Trials Resources and Publications Klinefelter Syndrome (KS): Condition Information Skip sharing on social media links Share this: Page Content What is KS? The term "Klinefelter (pronounced KLAHYN-fel-ter ) syndrome," ...

  2. Rett syndrome

    MedlinePLUS

    Rett syndrome occurs almost always in girls. It may be diagnosed as autism or cerebral palsy. Most Rett syndrome cases are due to a problem in the gene called MECP2. This gene is on the X chromosome. Females ...

  3. Metabolic Syndrome

    MedlinePLUS

    Metabolic syndrome is a group of conditions that put you at risk for heart disease and diabetes. These ... doctors agree on the definition or cause of metabolic syndrome. The cause might be insulin resistance. Insulin is ...

  4. Marfan Syndrome

    MedlinePLUS

    ... syndrome is a condition in which your body's connective tissue is abnormal. Connective tissue helps support all parts of your body. It ... and develops. Marfan syndrome most often affects the connective tissue of the heart and blood vessels, eyes, bones, ...

  5. Cushing syndrome

    MedlinePLUS

    Cushing syndrome is a disorder that occurs when your body has a high level of the hormone cortisol. ... The most common cause of Cushing syndrome is taking too much ... of this type of medicine. Glucocorticoids mimic the action ...

  6. Cushing's Syndrome

    MedlinePLUS

    Cushing's syndrome is a hormonal disorder. The cause is long-term exposure to too much cortisol, a hormone ... cause your body to make too much cortisol. Cushing's syndrome is rare. Some symptoms are Upper body obesity ...

  7. Angelman Syndrome

    MedlinePLUS

    ... heads, jerky movements, protruding tongues, and bouts of laughter." Infants with Angelman syndrome appear normal at birth, ... Is there any treatment? There is no specific therapy for Angelman syndrome. Medical therapy for seizures is ...

  8. Impairment of the Antibody-Dependent Phagocytic Function of PMNs through Regulation of the Fc?Rs Expression after Porcine Reproductive and Respiratory Syndrome Virus Infection

    PubMed Central

    Li, Peng; Jin, Qianyue; Liu, Yunchao; Bao, Dengke; Liu, Mingyang; Wang, Yinbiao; Zhang, Gaiping

    2013-01-01

    Porcine reproductive and respiratory syndrome (PRRS) is identified as one of the most important etiological agents in multifactorial respiratory disease of swine and can predispose pigs to secondary infections by other pathogens, usually bacteria. To understand the mechanism for an increased susceptibility to secondary bacterial infections, we investigated the antibody-dependent phagocytosis behaviour and killing ability of PMNs after infection by PRRSV strains BJ-4 or HN07-1. PMN’s antibody-dependent phagocytosis and their ability to kill E.coli were both noticeably decreased following PRRSV infection, in particular with the highly pathogenic strain HN07-1. As the change in this function of the PMNs may reflect a variation in the expression of Fc?Rs, the expression profiles of the activating and the inhibitory Fc?Rs were examined. We found that RNA expression of the inhibitory receptor Fc?RIIB was up-regulated post-infection, and this was greater after infection with the more virulent PRRSV strain HN07-1. The activating receptor Fc?RIIIA RNA expression was on the other hand inhibited to the same extent by both PRRSV strains. Neutralizing antibody titers post-infection by PRRSV strains BJ-4 or HN07-1 were also detected. All of the pigs in infection groups showed viraemia by the end of the study (56 DPI). These observations may help to understand the mechanism of increased susceptibility to secondary bacterial infections following PRRSV infection. PMID:23825597

  9. Impact of highly active antiretroviral therapy on oral manifestations of patients with human immunodeficiency virus/acquired immuno deficiency syndrome in South India

    PubMed Central

    Rao, K. V. S. Eswara; Chitturi, Ravi Teja; Kattappagari, Kiran Kumar; Kantheti, Lalith Prakash Chandra; Poosarla, Chandrasekhar; Baddam, Venkat Ramana Reddy

    2015-01-01

    Background: Human immunodeficiency virus (HIV) infection remains a global health problem, although the development of highly active antiretroviral therapy (HAART) has significantly modified the course of HIV disease into a manageable disease with improved quality-of-life mainly in the developed countries. Very few studies are available regarding effect of HAART on oral lesions in developing countries like India. Aims and Objectives: The aim was to document and compare oral lesions in HIV-seropositive patients before and after HAART. Materials and Methods: Oral manifestations were recorded in 320 HIV seropositive patients attending to the Voluntary Counseling and Confidential Testing Centre at the Government General Hospital, Guntur, before and after treating with HAART and the results were statistically analyzed using Student's t-test and Chi-square test. Results: Oral Candidiasis was significantly reduced in patients under HAART after 3 months. Furthermore, there was decreased incidence of periodontal diseases, but increased hyperpigmentation in patients undergoing HAART. Conclusion: The oral manifestations of HIV infection have changed due to the advent of HAART. Many opportunistic infections have resolved as a result of an improved immune system. Though the risk of hyperpigmentation in those with HAART has increased the prevalence of oral candidiasis and periodontal diseases were less in patients who had access to HAART. PMID:26392652

  10. Aase syndrome

    MedlinePLUS

    Aase-Smith syndrome; Hypoplastic anemia/Triphalangeal thumb syndrome ... Jones KL, ed. Aase syndrome. In: Smith's Recognizable Patterns Of Human Malformation. 6th ed. Saunders. 2005. Clinton C, Gazda HT. Diamond-Blackfan Anemia. 2009 Jun 25 [Updated 2013 Jul ...

  11. Down syndrome

    MedlinePLUS

    Down syndrome is a genetic condition in which a person has 47 chromosomes instead of the usual 46. ... In most cases, Down syndrome occurs when there is an extra copy of chromosome 21. This form of Down syndrome is called Trisomy 21. ...

  12. Velocardiofacial Syndrome

    ERIC Educational Resources Information Center

    Gothelf, Doron; Frisch, Amos; Michaelovsky, Elena; Weizman, Abraham; Shprintzen, Robert J.

    2009-01-01

    Velocardiofacial syndrome (VCFS), also known as DiGeorge, conotruncal anomaly face, and Cayler syndromes, is caused by a microdeletion in the long arm of Chromosome 22. We review the history of the syndrome from the first clinical reports almost half a century ago to the current intriguing molecular findings associating genes from the…

  13. Development of a fluorescent microsphere immunoassay for detection of antibodies against porcine reproductive and respiratory syndrome virus using oral fluid samples as an alternative to serum-based assays.

    PubMed

    Langenhorst, Robert J; Lawson, Steven; Kittawornrat, Apisit; Zimmerman, Jeffrey J; Sun, Zhi; Li, Yanhua; Christopher-Hennings, Jane; Nelson, Eric A; Fang, Ying

    2012-02-01

    For effective disease surveillance, rapid and sensitive assays are needed to detect antibodies developed in response to porcine reproductive and respiratory syndrome virus (PRRSV) infection. In this study, we developed a multiplexed fluorescent microsphere immunoassay (FMIA) for detection of PRRSV-specific antibodies in oral fluid and serum samples. Recombinant nucleocapsid protein (N) and nonstructural protein 7 (nsp7) from both PRRSV genotypes (type I and type II) were used as antigens and covalently coupled to Luminex fluorescent microspheres. Based on an evaluation of 488 oral fluid samples with known serostatus, the oral fluid-based FMIAs achieved >92% sensitivity and 91% specificity. For serum samples (n = 1,639), the FMIAs reached >98% sensitivity and 95% specificity. The assay was further employed to investigate the kinetics of the antibody response in infected pigs. In oral fluid, the N protein was more sensitive for the detection of early infection (7 and 14 days postinfection), but nsp7 detected a higher level and longer duration of antibody response (28 days postinfection). In serum, the antibodies specific to nsp7 and N proteins were detected as early as 7 days postinfection, and the responses lasted more than 202 days. This study provides a framework from which a more robust assay could be developed to profile the immune response to multiple PRRSV antigens in a single test. The development of oral fluid-based diagnostic tests will change the way we survey diseases in swine herds and improve our ability to cheaply and efficiently track PRRSV infections in both populations and individual animals. PMID:22155765

  14. Comparison of commercial real-time reverse transcription-PCR assays for reliable, early, and rapid detection of heterologous strains of porcine reproductive and respiratory syndrome virus in experimentally infected or noninfected boars by use of different sample types.

    PubMed

    Gerber, Priscilla F; O'Neill, Kevin; Owolodun, Olajide; Wang, Chong; Harmon, Karen; Zhang, Jianqiang; Halbur, Patrick G; Zhou, Lei; Meng, Xiang-Jin; Opriessnig, Tanja

    2013-02-01

    The aims of this study were to compare three commercial porcine reproductive and respiratory syndrome virus (PRRSV) real-time reverse transcription-PCR (RT-PCR) assays for detection of genetically diverse PRRSV isolates in serum, semen, blood swabs, and oral fluids collected from experimentally infected boars and to evaluate the effects of sample pooling. Six groups of three boars negative for PRRSV were each inoculated with one of six PRRSV isolates (sharing 55 to 99% nucleotide sequence identity in ORF5). Samples were collected on days -2, 1, 3, 5, 7, 14, and 21 postinoculation (p.i.) and tested by one of three commercially available real-time RT-PCR assays (VetMax from Applied Biosystems, Foster City, CA [abbreviated AB]; VetAlert from Tetracore, Rockville, MD [TC]; and AcuPig from AnDiaTec GmbH, Kornwestheim, Germany [AD]). At day 1 p.i., all assays detected at least one positive sample in each group. The highest detection rates were on days 3 and 5 p.i. Between days 1 and 7 p.i., serum samples had the highest detection rate (90%) with 100% agreement between tests, followed by blood swabs (kappa value of 0.97) and semen (kappa value of 0.80). Oral fluids had the lowest detection rates (AB, 55%; TC, 41%; AD, 46%) and the highest disagreement between kits (kappa value of 0.63). Pools of five samples did not reduce the detection rates if there was one positive sample with a large amount (cycle threshold, <30) of viral RNA in the pool. Serum and blood swab samples had shorter turnaround times for RNA extraction. The AB assay had a 1.6-times-shorter PCR time. In summary, serum and blood swabs had the best performance with highest detection rates and agreement between assays and the shortest turnaround times. PMID:23224085

  15. Molecular characterization of the porcine S100A6 gene and analysis of its expression in pigs infected with highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV).

    PubMed

    Zhou, Xiang; Wang, Peng; Michal, Jennifer J; Wang, Yan; Zhao, Jinhua; Jiang, Zhihua; Liu, Bang

    2015-08-01

    Our previous microarray study revealed that S100A6 was significantly upregulated in porcine alveolar macrophages (PAMs) infected with highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). In the present study, we cloned both cDNA and genomic DNA sequences of the gene. Transient transfection indicated that the porcine S100A6 protein was located in the nucleus and cytoplasm. Reverse transcription polymerase chain reaction (RT-PCR) revealed that the porcine S100A6 gene was highly expressed in the kidney and subcutaneous fat. Polyinosinic-polycytidylic acid [poly (I:C)] induced porcine S100A6 gene expression in PK-15 cells. Quantitative real-time PCR (Q-PCR) analysis further showed that the porcine S100A6 gene was upregulated in different cells and tissues of Tongcheng pigs infected with HP-PRRSV. Chromosome walking obtained the porcine S100A6 promoter region and then luciferase reporter assays confirmed its regulatory activities. We observed a putative NF-?B binding site in the core promoter region, which may explain the upregulation of porcine S100A6 in response to PRRSV. Transfection of NF-?B (p65 subunit) intensely induced the promoter activity of the porcine S100A6 gene, while an NF-?B inhibitor, pyrrolidine dithiocarbamate (PDTC), inhibited this activity. Furthermore, compared to its wild type, the promoter activity was significantly reduced when it contained a mutant NF-?B binding site. All these results provide a solid foundation to further investigate how S100A6 is involved in PRRSV infection. PMID:25480733

  16. Viruses of Haloarchaea

    PubMed Central

    Luk, Alison W. S.; Williams, Timothy J.; Erdmann, Susanne; Papke, R. Thane; Cavicchioli, Ricardo

    2014-01-01

    In hypersaline environments, haloarchaea (halophilic members of the Archaea) are the dominant organisms, and the viruses that infect them, haloarchaeoviruses are at least ten times more abundant. Since their discovery in 1974, described haloarchaeoviruses include head-tailed, pleomorphic, spherical and spindle-shaped morphologies, representing Myoviridae, Siphoviridae, Podoviridae, Pleolipoviridae, Sphaerolipoviridae and Fuselloviridae families. This review overviews current knowledge of haloarchaeoviruses, providing information about classification, morphotypes, macromolecules, life cycles, genetic manipulation and gene regulation, and host-virus responses. In so doing, the review incorporates knowledge from laboratory studies of isolated viruses, field-based studies of environmental samples, and both genomic and metagenomic analyses of haloarchaeoviruses. What emerges is that some haloarchaeoviruses possess unique morphological and life cycle properties, while others share features with other viruses (e.g., bacteriophages). Their interactions with hosts influence community structure and evolution of populations that exist in hypersaline environments as diverse as seawater evaporation ponds, to hot desert or Antarctic lakes. The discoveries of their wide-ranging and important roles in the ecology and evolution of hypersaline communities serves as a strong motivator for future investigations of both laboratory-model and environmental systems. PMID:25402735

  17. Ebolavirus and Haemorrhagic Syndrome

    PubMed Central

    Matua, Gerald A.; Van der Wal, Dirk M.; Locsin, Rozzano C.

    2015-01-01

    The Ebola virus is a highly virulent, single-stranded ribonucleic acid virus which affects both humans and apes and has fast become one of the world’s most feared pathogens. The virus induces acute fever and death, with haemorrhagic syndrome occurring in up to 90% of patients. The known species within the genus Ebolavirus are Bundibugyo, Sudan, Zaïre, Reston and Taï Forest. Although endemic in Africa, Ebola has caused worldwide anxiety due to media hype and concerns about its international spread, including through bioterrorism. The high fatality rate is attributed to unavailability of a standard treatment regimen or vaccine. The disease is frightening since it is characterised by rapid immune suppression and systemic inflammatory response, causing multi-organ and system failure, shock and often death. Currently, disease management is largely supportive, with containment efforts geared towards mitigating the spread of the virus. This review describes the classification, morphology, infective process, natural ecology, transmission, epidemic patterns, diagnosis, clinical features and immunology of Ebola, including management and epidemic containment strategies. PMID:26052448

  18. Inhibition of porcine reproductive and respiratory syndrome virus replication with exosome-transferred artificial microRNA targeting the 3' untranslated region.

    PubMed

    Zhu, Li; Bao, Liping; Zhang, Xinyu; Xia, Xiaoli; Sun, Huaichang

    2015-10-01

    Porcine reproductive and respiratory syndrome (PRRS) is an economically important swine disease. As part of the development of RNA interference (RNAi) strategy against the disease, in this study a recombinant adenovirus (rAd) expressing the artificial microRNA (amiRNA) targeting the 3' untranslated region (UTR) was used to investigate the exosome-mediated amiRNA transfer from different pig cell types to porcine alveolar macrophages (PAMs). Quantitative RT-PCR showed that the sequence-specific amiRNA was expressed in and secreted via exosomes from the rAd-transduced pig kidney cell line PK-15, PAM cell line 3D4/163, kidney fibroblast cells (PFCs) and endometrial endothelial cells (PEECs) with different secretion efficiencies. Fluorescent microscopy revealed that the dye-labeled amiRNA-containing exosomes of different cell origins were efficiently taken up by all of the five types of pig cells tested, including primary PAMs. Quantitative RT-PCR showed that the amiRNA-containing exosomes of different cell origins were taken up by primary PAMs in both time- and dose-dependent manners. Both quantitative RT-PCR and viral titration assays showed that the exosome-delivered amiRNA had potent anti-viral effects against three different PRRSV strains. These data suggest that the exosomes derived from pig cells could serve as an efficient miRNA transfer vehicle, and that the exosome-delivered amiRNA had potent anti-viral effects against different PRRSV strains. PMID:26238924

  19. Hepadna viruses

    SciTech Connect

    Robinson, W.; Koike, K.; Will, H.

    1987-01-01

    This book examines the molecular biology, disease pathogenesis, epidemiology, and clinical features of hepadna and other viruses with hepatic tropism and outlines future directions and approaches for their management. The volume's six sections provide a review of the various features, mechanisms, and functions of these viruses, ranging from hepadna virus replication and regulation of gene expression to the structure and function of hepadna-virus gene products.

  20. Characterization of the Uukuniemi virus group (Phlebovirus: Bunyaviridae): evidence for seven distinct species.

    PubMed

    Palacios, Gustavo; Savji, Nazir; Travassos da Rosa, Amelia; Guzman, Hilda; Yu, Xuejie; Desai, Aaloki; Rosen, Gail Emilia; Hutchison, Stephen; Lipkin, W Ian; Tesh, Robert

    2013-03-01

    Evolutionary insights into the phleboviruses are limited because of an imprecise classification scheme based on partial nucleotide sequences and scattered antigenic relationships. In this report, the serologic and phylogenetic relationships of the Uukuniemi group viruses and their relationships with other recently characterized tick-borne phleboviruses are described using full-length genome sequences. We propose that the viruses currently included in the Uukuniemi virus group be assigned to five different species as follows: Uukuniemi virus, EgAn 1825-61 virus, Fin V707 virus, Chizé virus, and Zaliv Terpenia virus would be classified into the Uukuniemi species; Murre virus, RML-105-105355 virus, and Sunday Canyon virus would be classified into a Murre virus species; and Grand Arbaud virus, Precarious Point virus, and Manawa virus would each be given individual species status. Although limited sequence similarity was detected between current members of the Uukuniemi group and Severe fever with thrombocytopenia syndrome virus (SFTSV) and Heartland virus, a clear serological reaction was observed between some of them, indicating that SFTSV and Heartland virus should be considered part of the Uukuniemi virus group. Moreover, based on the genomic diversity of the phleboviruses and given the low correlation observed between complement fixation titers and genetic distance, we propose a system for classification of the Bunyaviridae based on genetic as well as serological data. Finally, the recent descriptions of SFTSV and Heartland virus also indicate that the public health importance of the Uukuniemi group viruses must be reevaluated. PMID:23283959

  1. Immune responses of infants to infection with respiratory viruses and live attenuated respiratory virus candidate vaccines.

    PubMed

    Crowe, J E

    1998-01-01

    Respiratory viruses such as respiratory syncytial virus (RSV), the parainfluenza viruses (PIV), and the influenza viruses cause severe lower respiratory tract diseases in infants and children throughout the world. Experimental live attenuated vaccines for each of these viruses are being developed for intranasal administration in the first weeks or months of life. A variety of promising RSV, PIV-3, and influenza virus vaccine strains have been developed by classical biological methods, evaluated extensively in preclinical and clinical studies, and shown to be attenuated and genetically stable. The ongoing clinical evaluation of these vaccine candidates, coupled with recent major advances in the ability to develop genetically engineered viruses with specified mutations, may allow the rapid development of respiratory virus strains that possess ideal levels of replicative capacity and genetic stability in vivo. A major remaining obstacle to successful immunization of infants against respiratory virus associated disease may be the relatively poor immune response of very young infants to primary virus infection. This paper reviews the immune correlates of protection against disease caused by these viruses, immune responses of infants to naturally-acquired infection, and immune responses of infants to experimental infection with candidate vaccine viruses. PMID:9711783

  2. 9. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    9. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Photographer and date unknown VIEW BETWEEN PIERS 2 AND 3, LOOKING FROM WAREHOUSE ROOF - Brooklyn Army Supply Base, Pier 2, Brooklyn, Kings County, NY

  3. 17. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    17. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Signal Corps, USA, 1947 VIEW OF PIER 4 AND PIER 5-BROOKLYN ARMY BASE TERMINAL - Brooklyn Army Supply Base, Pier 4, Brooklyn, Kings County, NY

  4. 18. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Signal Corps, USA, 1933 VIEW OF EAST (FRONT) ELEVATION-PIER 4 - Brooklyn Army Supply Base, Pier 4, Brooklyn, Kings County, NY

  5. 6. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Topo-Metrics, Inc, 1992. Aerial view of the Brooklyn Army Terminal - Brooklyn Army Supply Base, Pier 2, Brooklyn, Kings County, NY

  6. 19. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    19. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Photographer and date unknown PIER 4 CONNECTING BRIDGE AND WAREHOUSE A - Brooklyn Army Supply Base, Pier 4, Brooklyn, Kings County, NY

  7. 10. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    10. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Signal Corps, USA, 1945 INTERIOR VIEW OF PIER SHED - Brooklyn Army Supply Base, Pier 2, Brooklyn, Kings County, NY

  8. 8. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    8. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Signal Corp, USA, 1946 VIEW OF PIERS 2, 3, AND 4-BROOKLYN ARMY BASE TERMINAL - Brooklyn Army Supply Base, Pier 2, Brooklyn, Kings County, NY

  9. 20. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    20. Photocopy of photograph (original in possession of NYC Economic Development Corp.) US Army photograph, 1949 VIEW SOUTH ELEVATION, OUTER END-PIER 4 - Brooklyn Army Supply Base, Pier 4, Brooklyn, Kings County, NY

  10. 7. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    7. Photocopy of photograph (original in possession of NYC Economic Development Corp.) Photographer unknown, circa 1983) OVERALL VIEW OF THE BROOKLYN ARMY TERMINAL, LOOKING NORTHEAST - Brooklyn Army Supply Base, Pier 2, Brooklyn, Kings County, NY

  11. 12. Photocopy of photograph (original in possession of NYC Economic ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    12. Photocopy of photograph (original in possession of NYC Economic Development Corp.) US Army Photograph, 1952 VIEW OF TEST HOLES BETWEEN PIERS - Brooklyn Army Supply Base, Pier 2, Brooklyn, Kings County, NY

  12. 18. Photocopy of photograph (original in possession of Chiricahua National ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. Photocopy of photograph (original in possession of Chiricahua National Monument), photographer unknown, c.1920's GROUP OF DUDES POSED IN FRONT OF SOUTH SIDE OF MAIN HOUSE - Faraway Ranch, Willcox, Cochise County, AZ

  13. 18. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    18. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Samuel H. Kneass, architect, January 1834. PLAN OF THE PRINCIPAL STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  14. 23. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    23. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. William Strickland, architect, circa December 1833. THIRD STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  15. 19. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    19. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Samuel H. Kneass, architect, January 1834. PLAN OF THE THIRD STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  16. 33. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    33. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Walter F. Price, architect, June 5, 1913. PROPOSED COVERED PASSAGES FOR THE PENNSYLVANIA HOSPITAL - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  17. 27. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    27. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. John C. Trautwine, architect and engineer, circa December 1833. NORTH FRONT and EAST & WEST FRONTS - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  18. 21. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    21. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. William Strickland, architect, circa December 1833. BASEMENT STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  19. 31. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    31. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Thomas U. Walter, architect, November 25, 1833. FRONT ELEVATION - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  20. 24. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    24. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. William Strickland, architect, circa December 1833. NORTH FRONT - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  1. 15. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    15. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. John Haviland, architect, December 1833. ELEVATION OF THE NORTH OR PRINCIPAL FRONT - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  2. 30. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    30. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Thomas U. Walter, architect, November 25, 1833. PLAN OF THIRD STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  3. 29. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    29. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Thomas U. Walter, architect, November 25, 1833. PLAN OF PRINCIPAL STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  4. 28. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    28. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Thomas U. Walter, architect, November 25, 1833. PLAN OF BASEMENT STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  5. 25. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    25. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. John C. Trautwine, architect and engineer, circa December 1833. PLAN OF THE BASEMENT - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  6. 16. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    16. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. John Haviland, architect, December 1833. LONGITUDINAL SECTION FROM EAST TO WEST - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  7. 26. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    26. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. John C. Trautwine, architect and engineer, circa December 1833. PLAN OF THE PRINCIPAL STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  8. 17. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    17. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Samuel H. Kneass, architect, January 1834. PLAN OF THE BASEMENT STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  9. 20. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    20. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. Samuel H. Kneass, architect, January 1834. PINE STREET FRONT - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  10. 14. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    14. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. John Haviland, architect, December 1833. PLAN OF THE PRINCIPAL FLOOR - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  11. 22. Photograph of architectural competition drawing; original in the possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    22. Photograph of architectural competition drawing; original in the possession of the Pennsylvania Hospital. William Strickland, architect, circa December 1833. SECOND STORY - Pennsylvania Hospital, Eighth & Ninth, Pine & Spruce Streets, Philadelphia, Philadelphia County, PA

  12. The epistemological significance of possession entering the DSM.

    PubMed

    Stephenson, Craig

    2015-09-01

    The discourse of the American Psychiatric Association's DSM reflects the inherently dialogic or contradictory nature of its stated mandate to demonstrate both 'nosological completeness' and cultural 'inclusiveness'. Psychiatry employs the dialogic discourse of the DSM in a one-sided, positivistic manner by identifying what it considers universal mental disease entities stripped of their cultural context. In 1992 the editors of the Diagnostic and Statistical Manual of Mental Disorders proposed to introduce possession into their revisions. A survey of the discussions about introducing 'possession' as a dissociative disorder to be listed in the DSM-IV indicates a missed epistemological break. Subsequently the editors of the DSM-5 politically 'recuperated' possession into its official discourse, without acknowledging the anarchic challenges that possession presents to psychiatry as a cultural practice. PMID:26254126

  13. 50 CFR 648.86 - NE Multispecies possession restrictions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...Regional Administrator projects that an in-season adjustment TAL trigger level for any small-mesh multispecies stock...reduction in the possession limit would be expected to prevent the TAL from being reached. (i) Red hake. If a...

  14. 1. Photographic copy of photograph, in possession of SCIP Office, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    1. Photographic copy of photograph, in possession of SCIP Office, COolidge, AZ. No date, circa 1935. Photographer unknown. COOLIDGE DIESEL PLANT - San Carlos Irrigation Project, Coolidge Diesel Plant, Off Highway 57, Coolidge, Pinal County, AZ

  15. 28. Photocopy of drawing (original in possession of Allegany County, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    28. Photocopy of drawing (original in possession of Allegany County, Cumberland, MD) FACTORY BUILDING, PLAN OF FLOOR B, MACHINERY LAYOUT, 1943, NOTE INFILLED COURTS - Kelly-Springfield Tire Plant, Factory Building, 701 Kelly Road, Cumberland, Allegany County, MD

  16. 27. Photocopy of drawing (original in possession of Allegany County, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    27. Photocopy of drawing (original in possession of Allegany County, Cumberland, MD) FACTORY BUILDING, PLAN OF FLOOR B, 1928, NOTE OPEN COURTS - Kelly-Springfield Tire Plant, Factory Building, 701 Kelly Road, Cumberland, Allegany County, MD

  17. 15. Photocopy of photograph (original print in possession of Earl ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    15. Photocopy of photograph (original print in possession of Earl & Donna Harris, 22802 Prospect Road, Cupertino, CA) 1914, photographer unknown VIEW OF HOUSE AFTER LANDSCAPING - Woodhills, Prospect Road, Cupertino, Santa Clara County, CA

  18. 14. Photocopy of photograph (original print in possession of Earl ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    14. Photocopy of photograph (original print in possession of Earl & Donna Harris, 22802 Prospect Road, Cupertino, CA) 1914, photographer unknown VIEW OF HOUSE BEFORE LANDSCAPING - Woodhills, Prospect Road, Cupertino, Santa Clara County, CA

  19. 15. Photographic copy of floor plan, not dated, in possession ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    15. Photographic copy of floor plan, not dated, in possession of Selfridge Base Museum, Mt. Clemens, Michigan. - Selfridge Field, Building No. 1050, Northwest corner of Doolittle Avenue & D Street; Harrison Township, Mount Clemens, Macomb County, MI

  20. 15. Photocopy of drawing (original in the possession of the ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    15. Photocopy of drawing (original in the possession of the Archives Collection, Syracuse Univerity) Photographer and date unknown PROPOSED DESIGN BY W. L. WOOLETT, ARCHITECT, SHOWING FRONT ELEVATION - Syracuse University, Hall of Languages, Syracuse University Campus, Syracuse, Onondaga County, NY